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For personal use only.Please do not reuse or reproducewithout the author’s permission.

Presented at the 2011 Stanford Mass Spectrometry Users’ Meeting

Healthy CarcinomaCirrhosis

C57B6 female micemice

(6 mo.)

ildWild type (WT)

Sod1 Sod1 Sod1

HCV‐S

Sod1 Sod1 Sod1

HCV‐FL

Sod1 Sod1 Sod1Sod1

‐/‐

Core E1 E2 p7 HCV-S

Core E1 E2 p7 NS3 NS4A NS4B NS5A NS5B HCV-FL

Treatments:

Control vs Alcohol Biochemical ProteomicsControl vs. Alcohol diets

analyses Proteomics

Treatments:

Control vs.

Biochemical analyses:

Liver enzyme activity; ProteomicsAlcohol

Liver enzyme activity;

Histopathology; etc…

Proteomics

Proteomics:

Treatments:

Control vs Alcohol

Biochemical analyses:

Detecting protein modification via

bisANS or disulfide Control vs. Alcohol

HistopathologybisANS or disulfide

labeling;

2D electrophoresis; Mass Spectrometry

15001400 -Control14001300

1200110010001000

900800

700600600500

400300200200100

0+/+ -/+ -/- +/+ -/+ -/- +/+ -/+ -/-

WT HCV-S HCV-FL

15001400 - Control14001300

120011001000

- Alcohol

900800

700600600500

400300200200100

0+/+ -/+ -/- +/+ -/+ -/- +/+ -/+ -/-

WT HCV-S HCV-FL

Control

Sod1 +/+ Sod1 -/+ Sod1 -/-Normal liver

Alcohol

Sod1 +/+ Sod1 -/+ Sod1 -/-Fatty liver

1 BisANS for protein conformational changes1. BisANS for protein conformational changes

1. 6‐IAF for protein modification

2-D gel electrophoresiAdd bisANS UV irradiation

sAdd bisANS U ad a o

& cross linkProtein extraction

ReducingAdd IAAProtein extraction

Reducing agent

Add IAA

2-D gel electrophoresi

sAdd 6-IAF

Total proteinvia

Coomassie blue

Hydrophobicityvia

bisANS

Disulfide bondvia

LC/MSD Trap XCT Ultra Agilent TechnologiesLC/MSD Trap XCT Ultra, Agilent Technologies

CytoplasmMetal-bindingPhosphoproteinAcetylationAlcohol catabolic processAlcohol catabolic processGlycolysis/GluconeogenesisMitochondrionMitochondrial lumen & matrixHomeostatic processAntioxidantOxidation reductionResponse to oxidative stressResponse to ROS

Response to H2O2Response to H2O2

Neg. regulation of apoptosis & death

DAVID Bioinformatics Resources, NIAID

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