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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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