Sulfate transport

Sulfate transport?

Renal sulfate transport occurs via a sulfate/bicarbonate exchange process.

!!

Misregulation of sulfate transport is present in a number of disease states including Alzheimer’s

disease.

Hofmeister seriesorganic anions > ClO4

- > SCN- > I- > NO3- > Br- > Cl- >

HCO3-> H2PO4

- > F-, SO42- > HPO4

2-

...orders anions in terms of their hydrophobicityF. Hofmeister, Arch. Exp. Patol. Phamakol. 1888, 24, 247.

Hydration energy

Anion ΔGh(X-) (kJmol-1)

NO3- -314

Cl- -338

HCO3- -388

SO42- -1103

B.A. Moyer et al. in Supramolecular Chemistry of Anions, Eds. A. Bianchi, K. Bowman-James and E. García-España, Wiley, 1997.

SO42-

phosphate buffer pH 7.2

Bicarbonate transport studies

HCO3-

pulse at 120s

Cl-

Chloride efflux promoted by 1-5 (2 % molar carrier to lipid) from unilamellar POPC vesicles loaded with 450 mM NaCl buffered to pH 7.2 with 20 mM sodium phosphate salts. The vesicles were dispersed in 162 mM Na2SO4 buffered to pH 7.2 with 20 mM sodium phosphate salts. At t = 120 s, a solution of NaHCO3 was added to give a 40 mM external concentration. At the end of the experiment, detergent was added to lyse the vesicles and calibrate the ISE to 100 % chloride efflux. Each point represents the

average of three trials. DMSO was used as control.

NH N HN

HNNH

HN

HN

R

RR

X

X

X1 X = O, R = Ph2 X = O, R = p-C6H4F3 X = O, R = C6F5

4 X = O, R = p-C6H4(CF3)2

5 X = O R=

CF3

CF3

SO42-


Sulfate transport?

HCO3-

pulse at 120s

Cl-

Chloride efflux promoted by 1-5 (2 % molar carrier to lipid) from unilamellar POPC vesicles loaded with 450 mM NaCl buffered to pH 7.2 with 20 mM sodium phosphate salts. The vesicles were dispersed in 162 mM Na2SO4 buffered to pH 7.2 with 20 mM sodium phosphate salts. At t = 120 s, a solution of NaHCO3 was added to give a 40 mM external concentration. At the end of the experiment, detergent was added to lyse the vesicles and calibrate the ISE to 100 % chloride efflux. Each point represents the

average of three trials. DMSO was used as control.

NH N HN

HNNH

HN

HN

R

RR

X

X

X6 X = S, R = Ph7 X = S, R = p-C6H4F8 X = S, R = C6F5

9 X = S, R = p-C6H4(CF3)2

10 X = S, R=

CF3

CF3

Lucigenin assay

Cl-

SO42- lucigenin Evidence for transmembrane transport of sulfate. Unilamellar POPC vesicles were loaded with 100

mM NaCl and 2 mM lucigenin buffered to pH 7.2 with 20 mM sodium phosphate salts and dispersed in a 100 mM NaCl solution (buffered to pH 7.2). At t = 30 s, a solution of the

appropriate anion was added (final concentration of 40 mM NaNO3 (red), 40 mM Na2SO4 (green) or 40 mM NaCl (blue)). At t = 60 s, a methanol solution of the putative transporter was added (2 %

molar carrier to lipid). At the end of the experiment (240 s), detergent was added to lyse the vesicles. Each point represents the average of three trials. A blank experiment was performed by

addition of Na2SO4, followed by addition of methanol; (a) compound 8.

N. Busschaert, M. Wenzel, M.E. Light, P.I. Hernadez, R. Perez-Tomas and P.A. Gale, J. Am. Chem. Soc. 2011, 133, 14136.

Cl-

NH N HN

HNNH

HN

HN

S

S

SF

F

F

F

F F

F

FF

F

FF

F

FF

Cyclic peptide-based receptors

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

N

8

NOO

NH N

O

HN OHN

NO

O

NHS NH

HN

HN S

NH

NHS

9

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

7

N. Busschaert, L.E. Karagiannidis, M. Wenzel, C.J.E. Haynes, N.J. Wells, P.G. Young, D. Makuc, J. Plavec, K.A. Jolliffe and P.A. Gale, Chem. Sci. 2014, DOI:10.1039/C3SC52006D.

Cl-/NO3- antiport

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

N

8

NOO

NH N

O

HN OHN

NO

O

NHS NH

HN

HN S

NH

NHS

9

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

7

NO3-Cl-


4% molar carrier to lipid

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

N

8

NOO

NH N

O

HN OHN

NO

O

NHS NH

HN

HN S

NH

NHS

9

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

7

Cl-/HCO3- antiportHCO3

- pulse at 120sCl-


SO42-


Lucigenin assay

NOO

NH N

O

HN OHN

NO

O

NHS NH

HN

HN S

NH

NHS

9

Cl-

SO42-

lucigenin

Cl-



Cl-/SO42- exchange

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

N

8

NOO

NH N

O

HN OHN

NO

O

NHS NH

HN

HN S

NH

NHS

9

NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

7

NH N HN

HNNH

HN

HN

R

RR

X

X

X

1 X=O R=C6H5 2 X=S R=C6H53 X=O R=C6F5 4 X=S R=C6F5 5 X=O R=m-C6H3(CF3)2 6 X=S R=m-C6H3(CF3)2



13C NMR assay for bicarbonate transport

O

-O

OH

Chloride pulse O

-O

OH

Cl-

+ Carrier

O

-O

OH

Cl-+ Mn2+

O

O-

OH

Cl-

Mn2+

NMR method: J.T. Davis, P.A. Gale, O.A. Okunola, P. Prados,

J.C. Iglesias-Sanchez, T. Torroba and R. Quesada Nature Chem. 2009, 1,

138-144. C&E News April 2009.

NH N HN

HNNH

HN

HN

R

RR

X

X

X 1 X = O, R = n-Bu2 X = O, R = Ph3 X = S, R = n-Bu4 X = S, R = Ph

33S NMR assay

NH N HN

HNNH

HN

HN

S

S

S NH N HN

HNNH

HN

HN

O

O

OF

F

F

F

F F

F

FF

F

FF

F

FF

NH N HN

HNNH

HN

HN

S

S

SF

F

F

F

F F

F

FF

F

FF

F

FF

33S NMR assay

NOO

NH N

O

HN OHN

NO

O

NHS NH

HN

HN S

NH

NHS

9NOO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

7

33S NMR assay

NH N HN

HNNH

HN

HN

S

S

S NH N HN

HNNH

HN

HN

O

O

OF

F

F

F

F F

F

FF

F

FF

F

FF

NH N HN

HNNH

HN

HN

S

S

SF

F

F

F

F F

F

FF

F

FF

F

FF

33S NMR assay

NOO

NH N

O

HN OHN

NO

O

NHS NH

HN

HN S

NH

NHS

9N

OO

NH N

O

HN OHN

NO

O

NH

S NH

HN

HNS

NH

NHS

7

Sulfate transport

Education

Transcript of Sulfate transport