181

Summary

Glutathione reductase (GR) is an enzyme that belongs to the oxidoreductase family of

flavoenzyme which catalysis the substrate (GSSG) into GSH in the presence of NADPH

(coenzyme) and FAD (cofactor). Significance of the enzyme is to maintain the homeostasis of

GSH:GSSG in cellular metabolism. As increasing GSSG, it is considered as oxidative stress

which effect on many cellular complications including lipid peroxidation, membrane

disintegration, protein de-folding and DNA breakage. Therefore, the GR is a catalyst for the

substrate-GSSG and it maintains GSSG in reduced state. Thus, the enzyme plays an important

role in Nostoc sp. which detoxifies the oxidative stress as the organism undergoes stressful

environment and protects the cellular metabolic activities namely photosynthesis, nitrogen

fixation, symbiotic relationships, detoxification of xenobiotic elements etc. Even though the

enzyme properties and mechanism were vastly reported in many organisms like E. coli, yeast,

parasites, plants and animals, it was not reported in Nostoc sp. Hence, GR was purified and

characterized from Nostoc sp. 701. However, sequencing of purified protein was found to be a

major task hence the similar sequences were retrieved from NCBI database. The sequences

were analyzed, characterized and finally undertaken for structure based theoretical analysis.

Ultimately, the following results were obtained,

Exponential growth of the organism Nostoc sp. 701 was attained in the culture medium

BG11 at 20th day. Protein and GR activity of the exponentially grown culture was

estimated to be 14.6 mg/ml and 0.109U/mg respectively. Ultimate purity of the GR was

obtained with specific activity of 149U/mg, 62% of protein recovery and 1230 fold

purification from Nostoc sp.701. The optimum pH, temperature and ionic strength of

the enzyme activity were found at pH 9.0, 55ºC and 50-150mM respectively. However,

the metal ions namely Hg2+

, Zn2+

, Cu2+

and Ag3+

influenced the enzyme activity.

Moreover, the molecular mass (Mr) of the subunit 53KDa was observed in SDS-PAGE

analysis. The kinetic analysis revealed that Km and Vmax of enzyme were estimated to be

0.058mM and 0.24µmol/min-1

ml-1

for GSSG; 0.0181mM and 0.192µmol/min-1

ml-1

for

NADPH. The Kcat was estimated to be 2.92×104M

-1 min

-1 and Kcat/Km of 5.04×10

8M

-1

min-1

for GSSG while Kcat and Kcat/Km of NADPH was estimated to be 2.34×104M

-1

min-1

and 12.92×108M

-1 min

-1 respectively.

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182

Sequence analysis of selected five sequences revealed Mr of 50KDa and other physico-

chemical properties of the enzyme was observed to be pI 5.52±0.41, GRAVY -0.068

±0.025, aliphatic index 91.8±3.37, total +ve charge amino acids (Arg and Lys) 45±3,

total -ve charge amino acids (Asp and Glu) 57±3, instability Index 25.41±6.24 and

extinction coefficient 30843±5862. Secondary structural properties of all five sequences

showed 32.45±1.16% of Alpha helix, 25.95±1.15% of Extended strand, 8.21±1.15% of

Beta turn and 33.37±1.13% of Random coil in Nostoc sp. 701.

Homology analysis revealed that Nostoc sp. PCC 7120 (BAB76667.1) shares 73% of

identity with other four Nostoc sp. sequences which was predicted from BLAST

analysis. Moreover, overall identity was 33.2% among the five sequences predicted

from ClustalW analysis. A novel conserved motif of TADKILIAVG ranging from 129

to 139 amino acids was found between the substrate and NAD binding domain as

observed in all five sequences.

The phylogenetic tree analysis revealed Nostoc sp. PCC 7120 (BAB76667.1), Nostoc

sp. PCC 7120 (CAA61856.1) and Nostoc punctiforme PCC 73102 (YP_001864581.1)

belongs to group-I whereas Nostoc azollae 0708 (ADI64813.1) group-II, and Nostoc

punctiforme PCC 73102 (ACC79657.1) observed as outgroup. Two novel motifs

LTPVAI and DSSIEGPGYSTM ranging from 329 to 335 and 381 to 392 respectively

were observed from evolutionary trace analysis.

Sub-cellular localization of the enzyme activity was observed in cytoplasm from the

amino acid sequence properties of GR. In addition to motif, architecture analysis

revealed GAGSGG, GGTCVIRGCVPK and GSGYIG fingerprints that predicted as

FAD, GSSG and NAD binding domain. Moreover, metabolic networking functions of

the enzyme were annotated.

The GR of Nostoc sp. PCC 7120 (BAB76667.1) sequence was modeled. The model was

isolated based on minimum DOPE score value of -102754.32 that revealed to be the

best model and the model was optimized therefore the energy minimum showed -

5071.827kJmol−1

. Finally, 91% of the amino acids were observed in most favored

region as 7.4% of amino acids were observed in allowed region and four outliers in the

Ramachandran plot from PROCHECK analysis.

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183

Docking analysis revealed that binding energy and ligand efficiency was observed to be

of -6.59kJ mol-1

and -0.16 per unit of polar surface area of the catalytic pocket

respectively. Inhibitory constant (Ki) was observed to be 14µM that could be the

minimum concentration of GSSG required to inhibit the enzyme. In addition to, the

non-bonded interaction energies intermolecular energy, vdw energy, electrostatic

energy, total energy and torsional energy were calculated.

Therefore, the enzyme complex was allowed to 5ns molecular dynamic simulation in

order to analyze potential, kinetic and total energies that was estimated to

1.33400e+06kJ mol-1

, -1.10874e+06kJ mol-1

and 2.25261e+05kJ mol-1

respectively. In

addition to, conformational changes of GR in explicit water model were analyzed

individually for FAD, NADPH and GSSG. Besides, hydrogen bonding pattern was

studied for GR along with ligand and substrate revealed that the complex marched

towards the equilibrium. As a result, the interaction profiles of the protein, ligand and

substrate revealed an evidence for the enzyme-substrate relationship as it required 1.5ns

for the substrate to be catalyzed.

* * *

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184

References

Alan B, Karam FA, The role of intracellular glutathione in inorganic mercury-induced toxicity

in neuroblastoma cells. Neurochem Res 2009;34:1677-1684.

Abd EM, Okada K, Goryo M, Kobayashi S, Oishi A, Yasuda J, Total glutathione and

glutathione reductase in bovine erythrocytes and liver biopsy. J Vet Med Sci

2008;70:861-864.

Agarkov AA, Popova TN, Semenikhina AV, Catalytic properties of glutathione reductase from

liver at norm and toxic hepatitis. Biochem (Moscow) Suppl Ser B: Biomed Chem

2009;3:172-176.

Akihiro KT, Hikaru S, Kunisuke T, Noriaki K, Kiyoshi T, Primary structure and properties of

glutathione reductase from Arabidopsis thaliana. Plant Cell Physiol 1993;34:1259-

1266.

Alcan NL, Tezcan EF, Sheep brain glutathione reductase: purification and general properties.

Elsevier Science Publishers B. V. (Biomedical Division) 1989;250:72-74.

Alfonso CG, Galisiieo EM, Antonio L, Rcena BJ, Barea JL, Horse-liver glutathione reductase:

purification and characterization. Inter J Biochem 1993;25:61-68.

Al-Senaidy AM, Purification and characterization of glutathione reductase from camel

(Camelus dromedaries) erythrocytes. Eur J Sci Res 2010;48:142-154.

Alton M, Selective modification of glutathione metabolism. Science 1983;220:472-477.

Alton M, Glutathione Metabolism and its selective modification. J Biol Chem 1988;263:17205-

17208.

Andrews P, Estimation of molecular size and molecular weights of biological compounds by

gel filtration. In Methods of Biochemical Analysis, lnterscience, New York. (Glikc, D.,

ed.). 1970;18:1-53.

Angela NK, Elisabeth DC, Ahmed AS, Lindsay LC, Jean D, Elias SJ, David LW, Thioredoxin

glutathione reductase from Schistosoma mansoni: An essential parasite enzyme and a

key drug target. PLoS Medicine 2007;4:1071-1086.

Angelucci F, Sayed AA, Williams AL, Boumis G, Brunori G, Dimastrogiovanni D, Miele AE,

Pauly F, Bellelli A, Inhibition of Schistosoma mansoni thioredoxin glutathione

reductase by auranofin: Structural and kinetic aspects. J Biol Chem 2009;284:28977-

28985.

Arias DG, Marquez VE, Beccaria AJ, Guerrero SA, Iglesias AA, Purification and

characterization of a glutathione reductase from Phaeodactylum tricornutum. Protist

2010;161:91-101.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

185

Arscott LD, Drake DM, Williams GH, Inactivation-Reactivation of Two-Electron Reduced

Escherichia coli Glutathione Reductase Involving a Dimer-Monomer Equilibrium.

Biochem 1989;28:3591-3598.

Asthana RK, Deepali, Tripathi DK, Arunima Srivastava A, Singh AP, Singh SP, Nath G,

Srivastava R, Srivastava BS, Isolation and identification of a new antibacterial entity

from the Antarctic cyanobacterium Nostoc CCC 537. J Appl Phycol 2009;21:81-88.

Babette KS, Brigit V, The stereospecificity of glutathione reductase for pyridine Nucleotides. J

Biol Chem 1960;235:209-212.

Badenhorst PW, Amory AM, Huckett BI, Light regulation of native and Escherichia coli

glutathione reductase in transgenic tobacco. J Plant Physiol W, L 1998;152:502-509.

Banu K, Kazim S, The interrelation of glutathione reductase, catalase, glutathione peroxidase,

superoxide dismutase, and glucose-6-phosphate in the pathogenesis of rheumatoid

arthritis. Clinic Rheumatol 2008;27:141-145.

Barea JL, Lee CY, Mouse-liver glutathione reductase purification, kinetics, and regulation. Eur

J Biochem 1979;98:487-499.

Basset RA, Friedl T, Mohr KI, Rybalka N, Martin W, High growth rate, photosynthesis rate

and increased hydrogen(ases) in manganese deprived cells of a newly isolated Nostoc-

like cyanobacterium (SAG 2306). Inter J Hydro Ener 2011;36:12200-12210.

Baxevanis AD, Ouellette BF, Bioinformatics-A practical guide to the analysis of genes and

proteins. Wiley Interscience, John wiley and sons publication. Second edition. 2001;83-

107.

Bashir K, Nagasaka S, Itai RN, Kobayashi T, Takahashi M, Nakanishi H, Mori S, Nishizawa

NK, Expression and enzyme activity of glutathione reductase is upregulated by Fe-

deficiency in graminaceous plants. Plant Mol Biol 2007;65:277-284.

Beesley CE, Smith RJ, Temple SJ, Lea PJ, Cloning and nucleotide sequence of the gene

encoding dinitrogenase reductase (nif H) from the cyanobacterium Nostoc 6720.

Biochimica Biophy Acta 1994;1219:548-550.

Berkholz DS, Faber HR, Savvides SN, Karplus PA, Catalytic cycle of human glutathione

reductase near 1 Å resolution. J Mol Biol 2008;382:371-384.

Bohme CC, Arscott LD, Becker K, Schirmer RH, Williams CH, Kinetic characterization of

glutathione reductase from the malarial parasite Plasmodium falciparum, comparison

with the human enzyme. J Biol Chem 2000;275:37317-37323.

Bhattacharya A, Tejero R, Montelione GT, Evaluating protein structures determined by

structural genomics consortia. Proteins 2007;66:778-95.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

186

Bhunia AK, Roy D, Banerjee SK, Carbaryl-induced effects on glutathione content, glutathione

reductase and superoxide dismutase activity of the cyanobacterium Nostoc muscorum.

Lett Appli Microbiol 1993;6:10-14.

Bradley KK, Bergman DK, Woods JP, Prevalence of American trypanosomiasis (Chagas

disease) among dogs in Oklahoma. J Am Vet Med Assoc 2000;217:1853-1857.

Buick R, When did oxygenic photosynthesis evolve? Phil Trans R Soc: B 2008;363:2731-2743.

Cakmak I, Activity of ascorbate-dependent H2O2- scavenging enzymes and leaf chlorosis are

enhanced in magnesium- and potassium-deficient leaves, but not in phosphorus-

deficient leaves. J Exp Bot 1994;45:1259-1266.

Campbell EL, Summers ML, Christman H, Martin ME, Meeks JC, Global gene expression

patterns of Nostoc punctiforme in steady-state dinitrogen-grown heterocyst-containing

cultures and at single time points during the differentiation of akinetes and hormogonia.

J bacterial 2007;189:5247-5256.

Can B, Erkmen GK, Dalmizrak O, Ogus HI, Ozer N, Purification and characterisation of rat

kidney glutathione reductase. Protein J 2010;29:250-256.

Candas M, Sohal RS, Radyuk SN, Klichko VI, Orr WC, Molecular organization of the

glutathione reductase gene in Drosophila melanogaster. Arch Biochem Biophy

1997;339:323-334.

Cardoso LA, Ferreira ST, Lima MH, Reductive inactivation of yeast glutathione reductase by

Fe(II) and NADPH. Comp Biochem Physiol Part A 2008;151:313-321.

Carlberg I, Mannervik B, Purification and characterization of the flavoenzyme glutathione

reductase from rat liver. J Biol Chem 1975;260:547-5480.

Carpenter EJ, Foster RA, Marine cyanobacterial symbioses. Biomed Life Sci, Cyanobacteria in

Symbiosis. 2003:11-17.

Chad K, Darryn W, The antioxidant role of glutathione and n-acetyl-cysteine supplements and

exercise-induced oxidative stress. J Inter Soc Sports Nutr 2005;2:38-44.

Chien LF, Vonshak A, Enzymatic antioxidant response to low-temperature acclimation in the

cyanobacterium Arthrospira platensis. J Appl Phycol 2011;23:887-894.

Chien TY, Chang DT, Chen CY, Weng YZ, Hsu CM, E1DS: catalytic site prediction based on

1D signatures of concurrent conservation. Nucleic Acids Res 2008;1:W291-W296.

Christine ES, Gordon RP, Susan TH, Lynette CE, Andrew MB, Dirk FM, Samuel L, Eric BG,

Harold F, Paul FE, Margie LC, Glutathione S-tranferase activity and glutathione S-

transferase expression in subjects with risk for colorectal cancer. Can Res

1995;55:2789-2793.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

187

Chwan YH, Yun-Yang C, Min-Yu Y, Sin-Yuan C, Shih-Chueh C, Ching Huei K, NaCl-

induced expression of glutathione reductase in roots of rice (Oryza sativa L.) seedlings

is mediated through hydrogen peroxide but not abscisic acid. Plant Soil 2009;320:103-

115.

Cohen SM, Olin KL, Feuer WJ, Hjelmeland L, Keen CL, Morse LS, Low glutathione reductase

and peroxidase activity in age-related macular degeneration. Br J Ophthalmol

1994;78:791-794.

Connell JP, Mullet JE, Pea Chloroplast glutathione reductase: purification and characterization.

Plant Physiol 1986;82:351-356.

Creissen GE, Edwards A, Enard C, Wellburn A, Mullineaux P, Molecular characterization of

glutathione reductase cDNAs from pea (Pisum sativum L.). Plant J 1992;2:129-131.

Cui J, Wakabayashi S, Wada K, Fukuyama K, Matsubara H, Isolation and sequence studies of

cysteinyl peptides from Spirulina glutathione reductase: comparison of active site

cysteine peptides with those of other flavoprotein disulfide oxidoreductases. J Biochem

1989;105:390-394.

David AD, Hanus FJ, Sterling AR, Harold JE, Purification, properties, and distribution of

ascorbate peroxidase in legume root nodules. Plant Physiol 1987;83:789-794.

Daniel RM, Dunn RV, Finney JL, Smith JC, The role of dynamics in enzyme activity. Ann Rev

Biophys Biomol Struct 2003;32:69-92.

Danyelle MT, Kenneth DT, Tapiero H, Dossier: Oxidative stress pathologies and antioxidants:

The importance of glutathione in human disease. Biomed Pharmacother 2003;57:145-

155.

Danielson UH, Jiang F, Hansson LO, Mannervik B, Probing the kinetic mechanism and

coenzyme specificity of glutathione reductase from the cyanobacterium Anabaena PCC

7120 by Redesign of the Pyridine-Nucleotide-Binding Site. Biochem1999;38:9254-

9263.

Deng Z, Hub Q, Lu F, Liu G, Hua Z, Colony development and physiological characterization

of the edible blue-green alga, Nostoc sphaeroides (Nostocaceae, Cyanophyta). Prog in

Nat Sci 2008;18:1475-1483.

Dere S, Gunes T, Sivaci R, Spectrophotometric determination of chlorophyll - A, B and total

carotenoid contents of some algae species using different solvents. Tr J Botany

1998;22:13-17.

Donough J, O‘Donovan, Julie PK, Toshiya T, Richard H, Xudong X, Charles VS, Stephen EW,

Gene transfer of mitochondrially targeted glutathione reductase protects H441 cells

from t-butyl hydroperoxides-induced oxidative stresses. Am J Respir Cell Mol Biol

1999;20:256-263.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

188

Dundas J, Ouyang Z, Tseng J, Binkowski A, Turpaz Y, Liang J, CASTp: computed atlas of

surface topography of proteins with structural and topographical mapping of

functionally annotated residues. Nucleic Acid Res 2006;34:W116-W118.

Durbin R, Eddy S, Krogh A, Mitchison G, Biological sequence analysis: probabilistic models

of proteins and nucleic acids. Camb uni press 2005:12-42.

Dutta D, Debojyoti D, Chaudhuri S, Bhattacharya SK, Review: Hydrogen production by

Cyanobacteria. Microb Cell Fact 2005;4:1-11.

Ekman M, Tollbäck P, Klint J, Bergman B, Protein expression profiles in an endosymbiotic

cyanobacterium revealed by a proteomic approach. MPMI 2006;19:1251-1261.

Elcock AH, Atomistic simulations of competition between substrates binding to an enzyme.

Biophys J 2002;82:2326-2332.

Elisabeth DC, Sandrine D, Christophe B, Babett S, Catharina CB, Andreas M, Louis M,

Christian S, Philippe G, Schirmer RH, Katja B, A prodrug form of a plasmodium

falciparum glutathione reductase inhibitor conjugated with a 4-Anilinoquinoline. J Med

Chem 2001;44:4268-4276.

Emanuelsson O, Brunak S, Heijne G, Nielsen H, Locating proteins in the cell using TargetP,

SignalP and related tools. Nat Protoc 2007;2:953-971.

Engle TE, Spears JW, Xi L, Edens FW, Dietary copper effects on lipid metabolism and

circulating catecholamine concentrations in finishing steers. J Anim Sci 2000;78:2737-

2744.

Erat M, Sakiroglu H, Ciftci M, Effects of some antibiotics on glutathione reductase from

bovine erythrocytes. Vet Med Czech 2003;48:305-312.

Ermakova EA, Zakhartchenko NL, Zuev YF, Brownian dynamics simulation of substrate

motion near active site of enzyme entrapped inside reverse micelle. Eur Biophys J

2010;39:1335-1341.

Ernest B, Effect of Flavin Compounds on Glutathione Reductase Activity: In Vivo and In Vitro

Studies. J Clini Invest 1969;48:1957-1966.

Eswar N, Eramian D, Webb B, Shen M, Sali A, Protein structure modeling with MODELLER,

Methods Mol Biol 2008;426:145-159.

Fan H, Mark AE, Zhu J, Honig B, Comparative study of generalized born models: protein

dynamics. PNAS 2005;102:6760-6764.

Fan H, Mark AE, Refinement of homology-based protein structures by molecular dynamics

simulation techniques. Protein Sci 2004;13:211-220.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

189

Farber PM, Becker K, Muller S, Schirmer RH, Franklin RM, Molecular cloning and

characterization of a putative glutathione reductase gene, the PfGR2 gene, from

Plasmodium falciparum. Eur J Biochem 1996;239:655-661.

Faixova Z, Faix, Influence of metal ions on ruminal enzyme activities. Acta Vet Brno

2002;71:451-455.

Fiore MF, Moon DH, Tsai SM, Lee H, Trevors JT, Miniprep DNA isolation from unicellular

and filamentous cyanobacteria. J Microbiol Method 2000;39:159-169.

Foyer CH, Souriau N, Perret S, Lelandais M, Over expression of glutathione reductase but not

glutathione synthetase leads to increases in antioxidant capacity and resistance to

photoinhibition in poplar trees. Plant Physiol 1995;109:1047-1057.

Foyer CH, Lelandais M, Galap C, Kunert KJ, Effects of elevated cytosolic glutathione

reductase activity on the cellular glutathione pool and photosynthesis in leaves under

normal and stress conditions. Plant Physiol 1991;97:863-872.

Galhanoa V, Laranjoa JG, Peixotob F, Exposure of the cyanobacterium Nostoc muscorum from

Portuguese rice fields to Molinate (Ordram): Effects on the antioxidant system and fatty

acid profile. Aqua Toxic 2011;101:367-376.

Gallo V, Schwarzer E, Rahlfs S, Schirmer RH, Zwieten R, Roos D, Arese P, Becker K,

Inherited Glutathione reductase deficiency and Plasmodium falciparum malaria-A Case

Study. PLoS ONE 2009;4:e7303

Gary C, Firmin J, Fryer M, Kular B, Leyland N, Reynolds H, Pastori G, Welburn F, Baker N,

Welburn A, Mullineaux PM, Elevated glutathione biosynthetic capacity in the

chloroplasts of transgenic tobacco paradoxically causes increased ocxidative stress.

Plant cell 1999;11:1277-1291.

Gaurav K, Banyal HS, Glutathione reductase and thioredoxin reductase: novel antioxidant

enzymes from Plasmodium berghei. Kor J Parasit 2009;47:421-424.

Geer LY, Domrachev M, Lipman DJ, Bryant SH, CDART: Protein Homology by domain

architecture. Genome Res 2002;12:1619-1623.

Geetha A, Lakshmi P, Jeyachristy SA, Surendran R, Level of oxidative stress in the red blood

cells of patients with liver cirrhosis. Ind J Med Res 2007;126:204-210.

George AK, Masao H, Arthur SS, Frank GE, William NV, Studies on chromated erythrocytes:

Effect of sodium chromate on erythrocyte glutathione reductase. J Clinic Invest

1964;43:323-331.

Gilberger T, Schirmer RH, Walter RD, Muller S, Deletion of the parasite-specific insertions

and mutation of the catalytic triad in glutathione reductase from chloroquine-sensitive

Plasmodium falciparum 3D7. Mol Biochem Parasit 2000;107:169-179.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

190

Giordanino MV, Strauch SM, Villafane VE, Helbling EW, Influence of temperature and UVR

on photosynthesis and morphology of four species of cyanobacteria. J Photochem

Photobiol B: Biology 2011;103:68-77.

Goktug U, Mustafa E, Mehmet C, Halis S, Ebubekir B, Purification and characterization of

glutathione Reductase from sheep liver. Tr J Vet Anim Sci 2005;29:1109-1116.

Graham N, Carolina MA, Jouanin L, Kunert KJ, Rennenberg H and Christine HF, Review

Article: Glutathione: biosynthesis, metabolism and relationship to stress tolerance

explored in transformed plants. J Experi Bot 1998;49:623-647.

Grau RU, Schirmer RH, Schirmer L, Siegel RL, Glutathione reductase from human

erythrocytes: amino-acid sequence of the structurally known fad-binding domain. Eur J

Biochern 1981;120:407-419.

Greer S, Perham RN, Glutathione reductase from Escherichia coli: cloning and sequence

analysis of the gene and relationship to other flavoprotein disulfide oxidoreductases.

Biochem 1986;25:2736-2742.

Griffith SM, Brewer TG, Steiner JJ, Thermal dependence of the apparent km of glutathione

reductase from three wetland grasses and maize. Ann Bot 2001;87:599-603.

Gul M, Kutay FZ, Temocin S, Cellular and clinical implications of glutathione. Ind J Exp Biol

2000;38:625-634.

Gutensohn W, Rodewald A, Haas B, Schulz P, Cleve H, Refined mapping of the gene for

glutathione reductase on human chromosome 8. Hum Genetics 1978;43:221-224.

Hakam N, Simon J, Molecular forms and thermal and kinetic properties of purified glutathione

reductase from two populations of barnyard grass (Echinochloa crus-galli (L.) Beauv.:

Poaceae) from contrasting climatic regions in North America. Can J Bot 2000;78:969-

980.

Halliwell B, Foyer CH, Properties and physiological function of a glutathione reductase

purified from spinach leaves by affinity chromatography. Planta 1978;139:9-17.

Han HS, Lee KD, Plant growth-promoting rhizobacteria: effect on antioxidant status,

photosynthesis, mineral uptake and growth of lettuce under soil salinity. Res J Agri Biol

Sci 2005;1:210-215.

Hans L, Dirk R, Ron W, Joap H, Familial deficiency of glutathione reductase in human blood

cells. Blood 1976;48:53-62.

Hargreaves P, Sheena Y, Land JM and Heales SJR, Glutathione deficiency in patients with

mitochondrial disease: Implications for pathogenesis and treatment. J Inherit Metab Dis

2005;28:81-88.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

191

Harkins DM, Brinkac LM, Nierman WC, Burkholderia mallei GB8 horse 4, EEP85169, J

Craig Venter Institute 2009.

Hausladen A, Alscher RC, Purification and characterization of glutathione reductase lsozymes

specific for the state of cold hardiness of red spruce. Plant Physiol 1994;105:205-213

He YY, Hader DP, Reactive oxygen species and UV-B: effect on cyanobacteria. Photochem

Photobiol Sci 2002;1:729-736.

Hercbergs A, Simoni F, Holtzman F, Bar-Am J, Brenner FC, Leith JT, Erythrocyte glutathione

and tumor response to chemotherapy. Lancet 1992;339:1074-1076.

Hikichi N, Pauline M, Hansza M, Tapi O, A molecular dynamics study of glutathione

reductase. J Mol Struct (T) 1995;335:243-254.

Hironori K, Shigeto M, Mieko N, Takehiro M and Kunisuke T, Gene cloning and expression of

cytosolic glutathione reductase in rice (Oryza Sativa L.). Plant Cell Physiol

1998;39:1269-1280.

Holden MT, Heather Z, Paillot R, Steward KF, Webb K, Ainslie F, Jourdan T, Bason NC,

Holroyd NE, Mungall K, Quail MA, Sanders M, Simmonds M, Willey D, Brooks K,

Aanensen DM, Spratt BG, Jolley KA, Maiden MC, Kehoe M, Chanter N, Bentley SD,

Robinson C, Maskell DJ, Parkhill J, Waller AS, Genomic evidence for the evolution of

Streptococcus equi: host restriction, increased virulence, and genetic exchange with

human pathogens. PLoS Pathol 2009;5:E1000346.

Hong C, Chao Y, Yang M, Cheng S, Cho S, Kao C, NaCl-induced expression of glutathione

reductase in roots of rice (Oryza sativa L.) seedlings is mediated through hydrogen

peroxide but not abscisic acid. Plant Soil 2009;320:103-115.

Hopkins GF, Dixon M, On glutathione-A thermostable oxidation-reduction system. J Biol

Chem 1921;15:527-563.

Hosikian A, Lim S, Halim R, Danquah MK, Chlorophyll extraction from microalgae: A review

on the process engineering aspects. Inter J Chem Eng 2010:1-11.

Hou WM, Amjad A Cheplinsky AB, Direct Submission, glutathione reductase [(Rattus

norvegicus)] Locus: AAB18132; Accession: U73174.1. Biochem Mol Biol 1996.

Hsu CM, Chen CY, Liu BJ. MAGIIC-PRO: detecting functional signatures by efficient

discovery of long patterns in protein sequences. Nucleic Acids Res 2006;34:W356-

W361.

Hua SJ, Sun ZR, Support vector machine approach for protein subcellular localization

prediction. Bioinfo 2001;17:721-728.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

192

Hyoshin L, Hye WS, Hyun LB, Dong PH, Won IC, Jinki J, Genomic cloning and

characterization of glutathione reductase gene from Brassica campestris var.

Pekinensis. Mol Cells 2001;13:245-251.

Ichiseki T, Ueda Y, Katsuda S, Kitamura K, Kaneuji A, Matsumoto T, Oxidative stress by

glutathione depletion induces osteonecrosis in rats. Rheumatol 2006;45:287-290.

Innis CA, Shi J, Blundell, Evolutionary trace analysis of TGF-β and related growth factors

implications for site-directed mutagenesis. Prot Eng 2000;13:839-847.

Iribarne F, Paulino M, Aguilera S, Murphy M, Tapia O, Docking and molecular dynamics

studies at trypanothione reductase and glutathione reductase active sites. J Mol Model

2002;8:173-183.

Jehonathan HP, Inna B, John T, Ping JL, Gurmit S, Eduard F, Brian CW, Androgen induces

adaptation to oxidative stress in prostate cancer: implications for treatment with

radiation therapy. Neoplasia 2007;9:68-80.

Jiang F, Hellman U, Sroga GE, Bergman B, Mannervik B, Cloning, sequencing, and regulation

of the glutathione reductase gene from the cyanobacterium Anabaena PCC 7120. J Biol

Chem 1995;270:22882-22889.

Johnson TJ, Kariyawasam S, Wannemuehler Y, Mangiamele P, Johnson SJ, Doetkott C,

Skyberg JA, Lynne AM, Johnson JR, Nolan LK, The genome sequence of avian

pathogenic Escherichia coli strain O1:K1:H7 shares strong similarities with human

extraintestinal pathogenic E. coli genomes. J Bacteriol 2007;189:3228-3236.

Joon JL, Ian WD, Jung HR, Isolation, expression, and regulation of the pgr11 gene encoding

glutathione reductase absolutely required for the growth of Schizosaccharomyces

pombe. J Biol Chem 1997;272:23042-23049.

Karni L, Stephen J, Elisha T, Glutathione reductase activity in heterocysts and vegetative cells

of the cyanobacterium Nostoc muscorum. Arch Microbiol 1984;140:215-217.

Karplus PA, Schulz GE, Substrate binding and catalysis by glutathione reductase as derived

from refined enzyme: substrate crystal structures at 2 Å resolution. J Mol Biol

1989;210:163-180.

Kay WW, Murfitt KC, The determination of blood glutathione. J Biol Chem 1959;74:203-208.

Kelner MJ, Montoya MA, Structural organization of the human glutathione reductase gene:

determination of correct cDNA sequence and identification of a mitochondrial leader

sequence. Biochem Biophys Res Commun 2000;269;366-368.

Kesharwani M, Ganguly B, Das A, Bandyopadhyay T, Differential binding of bispyridium

oxime drugs with acetylcholinesterase. Acta Pharmacologica Sinica 2010;31:313-328.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

193

Kidambi SP, Mahan JR, Matches AG, Purification and thermal dependence of glutathione

reductase from two forage legume species. Plant Physiol 1990;92:363-367

Kim IS, Shin SY, Kim YS, Kim HY, Yoon HS, Expression of a glutathione reductase from

Brassica rapa subsp. pekinensis enhanced cellular redox homeostasis by modulating

antioxidant proteins in Escherichia coli. Mol Cells 2009;28:479-487.

Kotopoulis S, Schommartz A, Postema M, Sonic cracking of blue-green algae. Applied

Acoustics 2009;70:1306-1312.

Kutser T, Quantitative detection of chlorophyll in cyanobacterial blooms by satellite remote

sensing. Limnol Oceano 2004;49:2179-2189.

Laemmli U, Cleavage of structural proteins during the assembly of the head of bacteriophage

T4. Nature 1970;227:680-685.

Lamotte F, Liaud NV, Duviau MP, Kobrehel K, Glutathione reductase in wheat grain. 1.

Isolation and characterization. J Agric Food Chem 2000;48:4978-4983.

Latifi A, Ruiz M, Zhang C, Oxidative stress in cyanobacteria. FEMS Microbiol Rev

2009;33:258-278.

Lascano HR, Casano LM, Melchiorre MN, Trippi VS, Biochemical and molecular

characterisation of wheat chloroplastic glutathione reductase. Biologia Plantarum

2001;44:509-516.

Lascano HR, Gomez LD, Casano LM, Trippi VM, Wheat chloroplastic glutathione reductase

activity is regulated by the combined effect of pH, NADPH and GSSG. Plant Cell

Physiol 1999;40:683-690.

Lawrence PK, Kittichotirat W, Bumgarner RE, McDermott JE, Herndon DR, Knowles DP,

Srikumaran S, Genome sequences of Mannheimia haemolytica serotype A2: ovine and

bovine isolates. J Bacteriol 2010;192:1167-1168.

Letunic I, Doerks T, Bork P, SMART 6: recent updates and new developments. Nucleic Acids

Res 2009;37:D229-232.

Liang J, Edelsbrunner H, Woodward C. Anatomy of protein pockets and cavities: measurement

of binding site geometry and implications for ligand design. Protein Sci 1998;7:1884-

1897.

Liberton M, Howard B, Heuser J, Roth R, Pakrasi H, Ultrastructure of the membrane systems

in the unicellular cyanobacterium Synechocystis sp. Strain PCC 6803. Protoplasma

2006;227:129-138.

Lindahl E, Hess B, Spoel DV, GROMACS 3.0: A package for molecular simulation and

trajectory analysis. J Mol Model 2001;7:306-317.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

194

Liu H, Wang H, Shenvi S, Hagen TM, Liu RM, Glutathione metabolism during aging and in

Alzheimer disease. Ann N Y Academic Sci 2004;1019:346-349.

Lopez CV, Garcia MC, Fernandez FG, Bustos CS, Chisti Y, Sevilla JM, Protein measurements

of microalgal and cyanobacterial biomass. Biore Tech 2010;101:7587-7591.

Lowry O, Rosebrough J, Farr A, Randall R, Protein measurement with the Folin-Phenol

reagents. J Biol Chem 1951;193:265-275.

Mackey D, Holt BF, Wiig A, Dangl JL. RIN4 interacts with Pseudomonas syringae type III

effector molecules and is required for RPM1-mediated resistance in Arabidopsis. Cell

2002;108:743-754.

Madamanchi NR, Anderson JV, Alscher RG, Cramer CL, Hess JL, Purification of multiple

forms of glutathione reductase from pea (Pisum sativum L.) seedlings and enzyme

levels in ozone-fumigated pea leaves. Plant Physiol 1992;100:138-145.

Marchler-Bauer A, Shennan L, Anderson JB, Chitsaz F, Derbyshire MK, Scott CD, Fong JH,

Geer LY, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Jackson JD, Zhaoxi K,

Lanczycki CJ, Fu L, Archler GH, Mullokandov M, Omelchenko MV, Robertson CL,

Song JS, Thanki N, Yamashita RA, Zhang D, Zhang N, Zheng C, Bryant SH, CDD: a

Conserved Domain Database for the functional annotation of proteins. Nucleic Acids

Res 2011;39:225-229.

Mariana M, German R, Victorio T, Roberto R, Lascano HR, Superoxide dismutase and

glutathione reductase overexpression in wheat protoplast: photooxidative stress

tolerance and changes in cellular redox state. Plant Growth Regul 2008;57:57-68.

Mathab SB, Sharada D, Hepatic glutathione reductase and riboflavin concentration in

experimental deficiency of thiamin and riboflavin in rats. J Nutri 1971;102:443-448.

Matsuda N, Motohashi K, Yoshimura H, Nozaki A, Inoue K, Ohmori M, Hisabori T, Anti-

oxidative stress system in cyanobacteria, significance of type II peroxiredoxin and the

role of 1-cys peroxiredoxin in synechocystis sp. Strain PCC 6803. J Biol Chem

2005;280:840-846.

Meeks JC, Elhai J, Thiel T, Potts M, Larimer F, Lamerdin J, Predki P, Atlas R, An overview of

the genome of Nostoc punctiforme, a multicellular, symbiotic cyanobacterium.

Photosyn Res 2001;70:85-106.

Meeks J, Castenholz R, Growth and photosynthesis in an extreme thermophile Synechococcus

lividus (Cyanophyta). Arch Mikrobiol 1971;78:25-41.

Meldrum NU, Tarr HL, The reduction of glutathione by the warburg-christian system. Biochem

1934;24:108-115.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

195

Mena RP, Dinglasan RR, Fayard

BF, Rodriguez JV, Caraballo MF, Ortiz AB, Coppens I,

Lorena MJ, Janse CJ, Serrano AE, Glutathione reductase-null malaria parasites have

normal blood stage growth but arrest during development in the mosquito. J Biol Chem

2010;285:27045-27056.

Mittl PR, Schulz GE, Structure of glutathione reductase from Escherichia coli at 1.86 Å

resolution: Comparison with the enzyme from human erythrocytes. Protein Sci

1994;3:799-809.

Mittl PR, Berry A, Scrutton NS, Perham RN, Schulz GE, Anatomy of an engineered NAD-

binding site. Protein Sci 1994;3:1504-1514.

Montero S, Arriaga D, Soler J, Purification and characterization of glutathione reductase from

mycelium of Phycomyces blakesleeanus. Biochirnica Biophysica Acta 1988;952:56-66.

Mortensen RA, The effect of diet on the glutathione content of erythrocytes. J Biol Chem 1953,

239-243.

Mu Q, Marta K, Cholewa JL, Weifei Z, Smart EJ, Sulistio MS, Asmis R, Macrophages

decreases atherosclerotic lesion formation in low-density lipoprotein receptor–deficient

mice. J Amer Heart Asso 2007;1375-1382.

Muller S, Becker K, Bergmann B, Schirmer RH, Walter RD, Plasmodium falciparum

glutathione reductase exhibits sequence similarities with the human host enzyme in the

core structure but differs at the ligand-binding sites. Mol Biochem Parasit 1995;74:11-

18.

Mullineaux P, Enard C, Hellens R, Creissen G, Characterization of a glutathione reductase

gene and its genetic locus from pea (Pisum sativum L.). Planta 1996;200:186-194.

Mustafa E, Lokman O, Leonardo MC, Yavuz D, Effect of Polyamines on Glutathione

Reductase Activity in Spinach. Z Naturforsch 2008;63:260-266.

Nagarkar S, Williams GA, Subramanian G, Saha SK, Cyanobacteria-dominated biofilms: A

high quality food resource for intertidal grazers. Hydrobiologia 2004;512:89-95.

Narayan K, Tiwari S, Pabbi S, Dhar WD, Biodiversity analysis of selected cyanobacteria. Curr

Sci 2006;91:947-951.

Natesan R, Shanmugasundaram P, Extracellular phosphate solubilization by the

cyanobacterium Anabaena ARM310. J Biosci 1989;14:203-208.

Nath J, Rebhun LI, Effects of caffeine and other methylxanthines on the development and

metabolism of sea urchin eggs. Involvement of NADP+ and glutathione. J Cell Biol

1976;68:440-450.

Nigel SS, Alan B, Richard NP, Engineering of an inter-subunit disulphide bridge in glutathione

reductase from Escherichia coli. Fed Eur Biochem Soc 1988;241:46-50.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

196

Ninomiya M, Hitoshi S, Yuji Y, Hiroyuki T, Mamoru K, Antioxidative activity and chemical

constituents of edible terrestrial alga Nostoc commune Vauch. Biosci, Biotech Biochem

2011;75:2175-2177.

Nishihara E, Kondo K, Parvez MM, Takahashi K, Watanabe K, Tanaka K, Role of 5-

aminolevulinic acid (ALA) on active oxygenscavenging system in NaCl-treated spinach

(Spinacia oleracea). J Plant Physiol 2003;160:1085-1091.

Norberto AS, David G, Determination of glutathione reductase activity in microgram samples

of tissue, quantitative histologic distribution of the enzyme in the rat adrenal and effect

of adrenocorticotropin. J Histo and Cytochem 1967;16:185-190.

Noriyuki Y, Hideyuki S, Yasuhiko I, Toshiaki K, Hiroshi K, Shimohama S, Akinori A,

Proteasome inhibition induces glutathione synthesis and protects cells from oxidative

stress relevance to parkinson disease. J Biol Chem 2007;282:4364–4372.

Nosratola DV, Xiu QW, Fariba O, Behdad R, Induction of oxidative stress by glutathione

depletion causes severe hypertension in normal rats. Hyper 2000;36:142-146.

Nuray U, Berivan T, Purification and kinetic properties of glutathione reductase from bovine

liver. Mol Cell Biochem 2007;303:45–51.

Ogus H, Ozer N, On the kinetics of human erythrocyte glutathione disulfide reductase: does the

enzyme really play ‗ping-pong‘? Tur J Biol 1999;23:143-151.

Lichtarge O, Bourne HR and Cohen FE, An evolutionary trace method defines binding surfaces

common to protein families. J Mol Biol 1996;257:342-358.

Ondrej Z, Dalibor H, Sona K, Vojtech A, Grace JC, Libuse T, Ales H, Jaromir H and Rene K,

An investigation of-glutathioneplatinum (II) interactions by means of the flow injection

analysis using glassy carbon electrode. Sensors 2007;7:1256-1270.

Pai EF, Schulz GE, The catalytic mechanism of glutathione reductase as derived from x-ray

diffraction analyses of reaction intermediates. J Biol Chem 1983;258:1752-1757.

Pai EF, Karplus PA, Schulz GE, Crystallographic analysis of the binding of NADPH, NADPH

fragments, and NADPH analogues to glutathione reductase. Biochem 1988;27:4465-

4474.

Pasquier C, Promponas VJ, Hamodrakas SJ, PRED-CLASS: Cascading neural networks for

generalized protein classification and genome-wide applications. Proteins 2001;44:361-

369.

Petegem FV, Vos D, Savvides S, Vergauwen B, Beeumen JV, Understanding nicotinamide

dinucleotide cofactor and substrate specificity in class I flavoprotein disulfide

oxidoreductases: crystallographic analysis of a glutathione amide reductase. J Mol Biol

2007;374:883-889.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

197

Peters GH, Frimurer TM, Andersen JN, Olsen OH, Molecular dynamics simulations of protein-

tyrosine phosphatase 1B. II substrate-enzyme interactions and dynamics. Biophy J

2000;78:2191-2200.

Pevsner J, Bioinformatics and functional genomics, Chapter 7: Molecular Phylogeny and

Evolution. Wiley Online Library 2003:260-266.

Philippis D, Faraloni R, Margheri C, Sili MC, Herdman C, Vincenzini M, Morphological and

biochemical characterization of the exocellular investments of polysaccharide-

producing Nostoc strains from the Pasteur Culture Collection. World J Microb

Biotechnol 2000;16:655-661.

Picaud T, Desbois A, Electrostatic control of the isoalloxazine environment in the two-electron

reduced states of yeast glutathione reductase. J Biol Chem 2002;277:31715-31721.

Piggott AM, Karuso P, Fluorometric assay for the determination of glutathione reductase

activity. Ann Chem 2007;79:8769-8773.

Pigiets VP, Conley RR, Purification of thioredoxin, thioredoxin reductase, and glutathione

reductase by affinity chromatography. J Biol Chem 1977;252:636743-636772.

Pilon-Smits EA, Zhu YL, Sears T, Terry N, Overexpression of glutathion reductase in Brassica

juncea: Effects on cadmium accumulation and tolerance. Physiol Plant 2000;110:455-

460.

Quevillon E, Silventoinen V, Pillai S, Harte N, Mulder N, Apweiler R, Lopez R, InterProScan:

protein domains identifier. Nucleic Acids Res 2005;33:W116-W120.

Racker E, Glutathione reductase from bakers‘ yeast and beef liver. J Biol Chem 1955;217:855-

865.

Rahman I, Aruna K, Saibal KB, Assay for quantitative determination of glutathione and

glutathione disulfide levels using enzymatic recycling method. Nature Prot

2007;1:3159-3165.

Rajinder SD, Drought stress, Enzymes of glutathione metabolism, oxidation injury, and protein

synthesis in Tortula Ruralis. Plant Physiol 1991;95:648-651.

Rania MA, Taha HM, Antimicrobial activity of cyanobacteria and green microalgae:

Evaluation of medium component by placket-burman design for antimicrobial activity

of Spirulina platensis. Glo J Biotech Biochem 2008;3:22-32.

Rendon JL, Hernandez GM, Unfolding kinetics of glutathione reductase from cyanobacterium

Spirulina maxima. Arch Biochem Biophy 2001;387:265-272.

Rescigno M, Perham R, Structure of the NADPH-binding motif of glutathione reductase:

efficiency determined by evolution. Biochem. 1994;33:5721-5727.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

198

Richard WB, Donald A, Reduced glutathione and glutathione disulfide: Free radicals and

antioxidant protocols. Methods Mol Biol 1998;108:347-352.

Rippka R, Deruelles J, Waterbury J, Herdman M, Stanier R, Generic assignments, strain

histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 1979;111:1-

61.

Rhonald CL, Lichtarge O, PyETV: A PyMOL Evolutionary trace viewer to analyze functional

site predictions in protein complexes. Bioinfo 2010:1-2.

Rogers LK, Tamura T, Rogers BJ, Welty SE, Hansen TN, Smith CV, Analyses of glutathione

reductase hypomorphic mice indicate a genetic knockout. Toxicol Sci 2004;82:367-373.

Roland S, John FK, Role of oxidative modifications in Atherosclerosis. Physiol Rev

2004;84:1381–1478.

Rollins MF, Heide DM, Weisend CM, Kundert JA, Comstock KM, Suvorova ES, Capecchi

ER, Merrill GF, Schmidt EE, Hepatocytes lacking thioredoxin reductase 1 have normal

replicative potential during development and regeneration. J Cell Sci 2010;123:2402-

2412.

Ruiz JM, Blumwald E, Salinity-induced glutathione synthesis in Brassica napus. Planta

2002;214:965-969.

Sarchizian I, Ardelean II, Axenic culture of a diazotrophic filamentous cyanobacterium isolated

from mesothermal sulphurous springs (obanul mare – mangalia). Rom J Biol-Plant Biol

2010;55:47-53.

Sarma GN, Savvides SN, Becker K, Schirmer M, Schirmer RH and Karplus PA, Glutathione

reductase of the malarial parasite Plasmodium falciparum: Crystal structure and

inhibitor development. J Mol Biol 2003;328:893-907.

Sato S, Nakamura Y, Kaneko T, Katoh T, Asamizu E, Tabata S, Structural analysis of

Arabidopsis thaliana chromosome 3. I. Sequence features of the regions of 4,504,864

bp covered by sixty P1 and TAC clones. DNA Res 2000;7:131-135.

Savvides SN, Karplus PA, Kinetics and crystallographic analysis of human glutathione

reductase in complex with a Xanthene inhibitor. J Biol Chem 1996;271:8101-8107.

Schaedle M, Bassham JM, Chloroplast glutathione reductase. Plant Physiol 1977:59:1011-

1012.

Schultz J, Milpetz F, Bork P, Ponting CP, SMART, a simple modular architecture research

tool: Identification of signaling domains. PNAS 1998;95:5857-5864.

Schuttelkopf AW, Aalten DM, PRODRG - A tool for high-throughput crystallography of

protein-ligand complexes. Acta Crystallogr 2004;60:1355-1363.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

199

Schutzendubel A, Polle A, Plant responses to abiotic stresses: Heavy metal-induced oxidative

stress and protection by mycorrhization. J Exp Bot 2002;53:1351-1365.

Serrano A, Joaquin R, Manuel L, Purification and properties of glutathione reductase from the

cyanobacterium Anabaena sp. Strain 7119. J Bacteriol 1984;158:317-324.

Seefeldt T, Zhao Y, Chen W, Raza AS, Carlson L, Herman J, Stoebner A, Hanson S, Foll R,

Guan X, Characterization of a novel dithiocarbamate glutathione reductase inhibitor and

its use as a tool to modulate intracellular glutathione. J Biol Chem 2009;284:2729-2737.

Seetharam B, Vasanth R, Changes in lens and erythrocyte glutathione reductase in response to

exogenous flavin adenine dinucleotide and liver riboflavin content of rat on riboflavin

deficient diet. Nutri Res 2006;7:1203-1208.

Sethubathi GV, Prabu VA, Antibacterial activity of cyanobacterial species from

adirampattinam coast, Southeast coast of palk bay. Curr Res J Biol Sci 2010;2:24-26.

Sharmila U, Subramanya U, Krishnamohan S, Vanajakshamma K, Mamatha K, Mathias S,

Oxidant-antioxidant status in colorectal cancer patients before and after treatment. Ind J

Clini Biochem 2004;19:80-83.

Sheweita SA, Heavy metal-induced changes in the glutathione levels and glutathione

reductase/glutathione S-transferase activities in the liver of male mice. Inter J Toxicol

1998;17: 383-392.

Sheehan D, Meade G, Chemical modulation of chemotherapy resistance in cultured

oesophageal carcinoma cells. Chemoprotection, Biochem Soc Trans 2000;28:27-32.

Shu DF, Wang LY, Duan M, Deng YS, Meng QW, Antisense-mediated depletion of tomato

chloroplast glutathione reductase enhances susceptibility to chilling stress. Plant Physiol

Biochem 2011;49:1228-1237.

Shyam SB, Kaushik D, Stephanie DS Xiang F, John DR, James PK, Glutathione oxidation and

mitochondrial depolarization as mechanisms of nordihydroguaiaretic acid-induced

apoptosis in lipoxigenase-deficient FL5.12 cells. Toxicol Sci 2000;53:77-83.

Siegel RL, Arscott LD, Janas AS, Schirmer RH, Williams CH. Role of active site tyrosine

residues in catalysis by human glutathione reductase. Biochem 1998;37:13968-13977.

Siegel RL, Muller JG, Lottspeich F, Schirmer RH, Glutathione reductase and glutamate

dehydrogenase of Plasmodium fulcipurum, the causative agent of tropical malaria. Eur J

Biochem 1996;235:345-350.

Siegel LK, Inhoff O, Parasite-specific trypanothione reductaseas a drug target molecule.

Parasitol Res 2003;90:S77–S85.

Silverthorn, Unglaub D, Human Physiology: An Integrated Approach. Addison-Wesley

Publication 2004:83-87.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

200

Smith IK, Vierheller TL, Thorne CA. Assay of glutathione reductase in crude tissue

homogenates using 5,5'-dithiobis(2-nitrobenzoic acid). Ann Biochem 1988;175:408-

413.

Stevens RG, Creissen GP, Mullineaux PM, Cloning and characterisation of a cytosolic

glutathione reductase cDNA from pea (Pisum sativum L.) and its expression in response

to stress. Plant Mol Biol 1997;35:641–654.

Stohs SJ, Rashidy FH, Lawson T, Kobayashi RH, Wulf BG , Potter JF, Changes in glutathione

and glutathione metabolizing enzymes in human erythrocytes and lymphocytes as a

function of age of donor. Age 1984;7:3-7.

Surapaneni KM, Vishnupriya V, Altered serum total sialic acid, lipid peroxidation,

ceruloplasmin and glutathione reductase levels in patients with carcinoma of prostate. J

Clinic Diag Res 2009;3:1483-1485.

Sven W, Kerstin W, Georg N, Regulation of antioxidant and oxidant enzymes in vascular cells

and implications for vascular disease. Medicine 2006;8:69-78.

Svircev Z, Simin DC, Simeunovic J, Karaman M, Stojanovic D, Antibacterial, antifungal and

cytotoxic activity of terrestrial cyanobacterial strains from Serbia. Sci China Series C:

Life Sci 2008;51:941-947.

Tamura T, McMicken HW, Smith CV, Hansen TN, Gene structure for mouse glutathione

reductase, including a putative mitochondrial targeting signal. Biochem Biophys Res

Commun 1997;237:419-422.

Tanaka K, Saji H, Kondo N, Immunological properties of spinach glutathione reductase and

inductive biosynthesis of the enzyme with ozone. Plant Cell Physiol 1988;29:637-642.

Tandogan B, Ulusu NN, The inhibition kinetics of yeast glutathione reductase by some metal

ions. J Enzyme Inhib Med Chem 2007;22:489-495.

Tandogan B, Ulusu N, Purification and kinetics of bovine kidney cortex glutathione reductase.

Protein Pept Lett 2010;17:667-674.

Tang H, Wang X, Bowers JE, Ming R, Alam M, Paterson AH, Unraveling ancient hexaploidy

through multiply-aligned angiosperm gene maps. Genome Res 2008;18:1944-1954.

Taniguchi N, From the γ-glutamyl cycle to the glycan cycle: a road with many turns and

pleasant surprises. J Biol Chem 2009;284:34469–34478.

Takeda T, Ishikawa T, Higeoka SS, Hirayam S, Mitsunaga T, Purification and characterization

of glutathione reductase from chlamydomonas reinhardtii. J Gen Microbiol

1993;139:2233-2238.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

201

Tekman B, Ozdemir H, Senturk M, Ciftci M, Purification and characterization of glutathione

reductase from rainbow trout (Oncorhynchus mykiss) liver and inhibition effects of

metal ions on enzyme activity. Comp Biochem Physiol Part C: Toxicol Pharma

2008;148:117-121.

Thacker R, Diaz C, Rutzler K, Erwin P, Kimble S, Pierce M, Dillard S, Phylogenetic

relationships among the filamentous cyanobacterial symbionts of Caribbean sponges

and a comparison of photosynthetic production between sponges hosting filamentous

and unicellular cyanobacteria. Por Res: Biodiversity Inno Sust 2007;621-626.

Theodore WR, Albert LL, Glutathione reductase of animal tissues. Biochem Nutr Found

1951:119-130.

Toshi K, Radhey S, Praveen V, Shoba S, Kumria MML, Sharma PC, Som NS, Glutathione

metabolism in rats exposed to highfluoride water and effect of Spirulina treatment.

Fluoride res rep 2001;34:132-138.

Torres LL, Quaglio NB, Souza GT, Garcia RT, Dati LM, Moreira WL, Loureiro AP, Talarico

JN, Smid J, Porto CS, Bottino CM, Nitrini R, Barros SB, Camarini R, Marcourakis T.

Peripheral oxidative stress biomarkers in mild cognitive impairment and Alzheimer's

disease. J Alzheim Dis 2011;26:59-68.

Treska V, Kobr J, Hasman D, Racek J, Trefil L, Reischig T, Hes O, Kuntscher V, Molacek J,

Treska I, Ischemia-reperfusion injury in kidney transplantation from non-heart-beating

donor –do antioxidants or anti-inflammatory drugs play any role? Exper Study Bratisl

Lek Listy 2007;110:133-136.

Turner L, Pollock C, Edmeades G, Thermal kinetics of glutathione reductase and their relation

to thermotolerance in diverse cultivars of maize. Ann Bot 1994;74:245-250.

Ulusu G, Erat M, Ciftci M, Sakiroglu H, Bakan E, Purification and characterization of

glutathione reductase from sheep liver. Turk J Vet Anim Sci 2005;29:1109-1117.

Ulusu. N, Tandogan B, Purification and kinetic properties of glutathione reductase from bovine

liver. Mol Cell Biochem 2008;303:45-51.

Vaishampayan A, Sinha RP, Hader P, Dey T, Gupta AK, Bhan U, Rao AL, Cyanobacterial

biofertilizers in rice agriculture. Bot Rev 2001;67: 453-516.

Valentina G, Evelin S, Stefan R, Heiner S, Rob van Z, Dirk R, Paolo A, Katja B, Inherited

Glutathione Reductase Deficiency and Plasmodium falciparum Malaria—A Case

Study. PLoS ONE 2009;4:e7303 (1-9).

Vijayaraj N, Elasri MO, Structure and function predictions of the Msa protein in

Staphylococcus aureus. BMC Bioinfo 2007;8:S5(1-9).

Wade RC, Simulation of enzyme-substrate interactions: The diffusional encounter step. Acta

Biochimica Polonica 1995;42:419-426.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

202

Wallen JR, Paige C, Mallett TC, Karplus PA, Claiborne A, Pyridine nucleotide complexes with

Bacillus anthracis coenzyme A-disulfide reductase: A structural analysis of dual

NAD(P)H specificity. Biochem 2008;47:5182–5193.

Wang G, Hu C, Li D, Zhang D, Li X, Chen K, Liu Y, The response of antioxidant systems in

Nostoc sphaeroides against UV-B radiation and the protective effects of exogenous

antioxidants. Ad Space Res 2007;39:1034-1042.

Wen J, Vyatkina G, Garg N, Oxidative damage during chagasic cardiomyopathy development:

Role of mitochondrial oxidant release and inefficient antioxidant defense. Free radical

Biol Med 2004;37:1821-1833.

Wessel D, Flugge U, A method for the quantitative recovery of protein in dilute solution in the

presence of detergents and lipids. Ann Biochem 1984;138:141-143.

Whitton B, Potts M, The ecology of cyanobacteria: Their diversity in time and space. Springer

ISBN 0792347358, 2000:61-81.

Wilkins AD, Lua R, Erdin S, Ward RM, Lichtarge O, Sequence and structure continuity of

evolutionary importance improves protein functional site discovery and annotation.

Protein Sci 2010;19:1296-1311.

Willmore W, Storey K, Purification and properties of glutathione reductase from liver of the

anoxia tolerant turtle Trachemys scripta elegans. Mol Cell Biochem 2007;297:139-149.

Wingsle G, Purification and characterization of glutathione reductase from scots pine needles.

Physiologia Plantarum 1989;76:24-30.

Wolfgang J, Georg ES, The binding of the retro- analogue of glutathione disulfide to

glutathione reductase. J Biol Chem 1990;265:10443-10445.

Wu G, Fang YZ, Yang S, Lupton JR, Turner ND, Glutathione metabolism and its implications

for health. J Nutri 2004;134:489-492.

Xu Y, Jacques PHC, Jiang H, Silman I, Sussman JL, Weik M, Induced-fit or preexisting

equilibrium dynamics? Lessons from protein crystallography and MD simulations on

acetylcholinesterase and implications for structure-based drug design. Protein Sci

2008;17:601-605.

Yamamoto Y, Kamio Y, Higuchi M, Cloning, nucleotide sequence, and disruption of

Streptococcus mutans glutathione reductase gene (gor). Biosci Biotech Biochem

1999;63:1056-1562.

Yang L, Song G, Jernigan RL, how well can we understand large-scale protein motions using

normal modes of elastic network models? Biophy J 2007;93:920-929.

Please purchase PDF Split-Merge on www.verypdf.com to remove this watermark.

203

Yannarelli YG, Alvarez AN, Cruz DM, Tomaro ML, Glutathione reductase activity and

isoforms in leaves and roots of wheat plants subjected to cadmium stress. Phytochem

2007;68:505-512.

Yoshihito Y, Kouichi AT, Regulatory mechanism of glutathione reductase activity in human

red cells. Blood 1974;43:99-109.

Yu NY, Wagner JR, Laird MR, Melli G, Rey S, Lo R, Dao P, Sahinalp SC, Ester M, Foster LJ,

Brinkman FS. PSORTb 3.0: Improved protein subcellular localization prediction with

refined localization subcategories and predictive capabilities for all prokaryotes. Bioinfo

2010;26:1608-1615.

Yu J, Zhou C, Crystal structure of glutathione reductase Glr1 from the yeast Saccharomyces

cerevisiae. Proteins: Struct Funct Bioinfo 2007;68:972-979.

Yusong Y, Ji-Zhong C, Sharad SS, Manjit S, Utpal P, Sanjay A, Yogesh CA, Role of

glutathione S- transferase in protection against lipid peroxidation. J Biol Chem

2001;276:19220-19230.

Zdobnov EM, Apweiler R, InterProScan-An integration platform for the signature-recognition

methods in InterPro. Bioinfo 2001;17:847-848.

Zhang P, Kharchenko V, Dalgarno A, Matsumi Y, Nakayama T, Takahashi K, Zhang, Reply:

Phys Rev Lett 2009;103:159302.

Zhang J, Kirkham MB, Antioxidant responses to drought in sunflower and sorghum seedlings.

New Phytologist 1996;132:361-373.

Zhao Y, Seefeldt T, Chen W, Carlson L, Stoebner A, Hanson S, Foll R, Matthees D, Palakurthi

S, Guan X, Increase in thiol oxidative stress via glutathione reductase inhibition as a

novel approach to enhance cancer sensitivity to x-ray irradiation. Free radical Biol Med

2009;47:176-183.

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204

Webography

http://prosite.expasy.org/PS00076

http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.8.1.7

http://www.chem.qmul.ac.uk/iubmb/

http://microbewiki.kenyon.edu/index.php/Nostoc,

http://en.wikipedia.org/wiki/Nostoc

http://bioweb.uwlax.edu/bio203/s2008/kaisersa_jose/Classification.html

http://expasy.org/tools/

http://web.expasy.org/protparam/

http://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html

http://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE=Proteins

http://www.genome.jp/tools/clustalw/

(http://mammoth.bcm.tmc.edu/uet/

http://mordred.bioc.cam.ac.uk/~jiye/evoltrace/evoltrace.html

http://linux1.softberry.com/berry.phtml?topic=pcompb&group=programs&subgroup=proloc

http://www.psort.org/psortb/

http://www.cbs.dtu.dk/services/TargetP/

http://sunflower.kuicr.kyoto-u.ac.jp/~smatsuda/slplocal.html

http://www.ebi.ac.uk/Tools/ppsearch/index.html

http://www.ncbi.nlm.nih.gov/Structure/lexington/lexington.cgi

http://meme.nbcr.net/meme4_6_0/cgi-bin/meme.cgi)

http://smart.embl-heidelberg.de/

http://www.ebi.ac.uk/Tools/InterProScan/

http://athina.biol.uoa.gr/PRED-CLASS/input.html

http://salilab.org/modeller/

http://spdbv.vital-it.ch/energy_tut.html

http://psvs-1_4-dev.nesg.org/

http://e1ds.csbb.ntu.edu.tw/

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205

http://sts.bioengr.uic.edu/castp/calculation.php

http://www.acdlabs.com/download/

www.gromacs.org

http://davapc1.bioch.dundee.ac.uk/prodrg/

http://plasma-gate.weizmann.ac.il/Grace/

http://urchin.nidcr.nih.gov/cgi-bin/blastREF.plx?tig=GLEAN3_25989

http://www.uniprot.org/uniprot/Q2RWK9

http://www.ncbi.nlm.nih.gov/Structure/cdd/cddsrv.cgi?uid=199420

http://www.ebi.ac.uk/interpro/IEntry?ac=IPR013027

http://salilab.org/modeller/methenz/andras/node16.html

http://www.ebi.ac.uk/thornton-srv/databases/cgi-bin/CSA/CSA_Show_EC_List.pl

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206

Publication

Ithayaraja M, Lakshmi PTV, Rajendhran J, and Gunasekaran P. Enzyme purification and

kinetics of glutathione reductase from Nostoc Sp. Strain 701. Under review by Brazilian

Journal of Microbiology.(Manuscript No. BJM-5409-72058-366789-2-SM).

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