E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.
-
Upload
jesse-adams -
Category
Documents
-
view
219 -
download
1
description
Transcript of E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.
![Page 1: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/1.jpg)
EXPLORING PROTEINS AND PROTEOMES
![Page 2: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/2.jpg)
GENOMES AND PROTEOMES
![Page 3: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/3.jpg)
OBTAINING THE PROTEIN: CELL LYSIS AND SEPARATION
![Page 4: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/4.jpg)
PROTEIN PURIFICATION TECHNIQUES Salting out
Dialysis
Gel Filtration Chromatography
Ion-exchange Chromatography
Affinity Chromatography
HPLC
Gel Electrophoresis
Isoelectric Focusing
2D Gel Electrophoresis
Ultracentrifugation
![Page 5: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/5.jpg)
SALTING OUT
![Page 6: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/6.jpg)
DIALYSIS
![Page 7: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/7.jpg)
GEL FILTRATION CHROMATOGRAPHY
![Page 8: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/8.jpg)
ION-EXCHANGE CHROMATOGRAPHY
![Page 9: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/9.jpg)
AFFINITY CHROMATOGRAPHY
![Page 10: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/10.jpg)
ELISA
![Page 11: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/11.jpg)
HPLC
![Page 12: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/12.jpg)
GEL ELECTROPHORESIS
![Page 13: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/13.jpg)
ISOELECTRIC FOCUSING
![Page 14: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/14.jpg)
2D GEL ELECTROPHORESIS
![Page 15: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/15.jpg)
ULTRACENTRIFUGATION
![Page 16: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/16.jpg)
STRUCTURE DETERMINATION
![Page 17: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/17.jpg)
AMINO ACID SEQUENCING: EDMAN DEGRADATION
![Page 18: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/18.jpg)
BROAD CLASSIFICATION OF PROTEASE Serine Proteases
A serine residue acts as a nucleophile in the active site, facilitating the reaction
Operates via the catalytic triad His, Ser- Asp
Cystein Proteases Cys as the nucleophile and activated by a nearby basic aa Papain is an example
Aspartate Proteases Two Asp residues cleaves the peptide bond by activating a
water molecule
Metalloprotease A metallic Zn or Co is involved in the catalytic mechanism
![Page 19: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/19.jpg)
EXAMPLES OF PROTEASES Trypsin – carboxyl side of Lys and Arg
Chymotrypsin – carboxyl side of bulky hydrophobic aa
Elastase – carboxyl side of small hydrophobic aa
Clostripain – carboxyl side of Arg
Thrombin – serine protease with specific cleavage site: Leu-Val-Pro-Arg—Gly-Ser
Carboxypeptidase A – amino side of C terminal residues of aromatic or aliphatic aa An exopeptidase
![Page 20: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/20.jpg)
MASS SPECTROMETRY
![Page 21: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/21.jpg)
MALDI-TOF MS
![Page 22: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/22.jpg)
ELECTROSPRAY IONIZATION
![Page 23: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/23.jpg)
X-RAY CRYSTALLOGRAPHY
![Page 24: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/24.jpg)
X-RAY DIFFRACTION PATTERN OF MEVALONATE KINASE
![Page 25: E XPLORING P ROTEINS AND P ROTEOMES. G ENOMES AND P ROTEOMES.](https://reader034.fdocuments.net/reader034/viewer/2022051318/5a4d1ad07f8b9ab059971377/html5/thumbnails/25.jpg)
NMR SPECTROSCOPY