for do not reuse or reproduce - stanford university...for personal use only. please do not reuse or...
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For personal use only.Please do not reuse or reproducewithout the author’s permission.
2
Presented at the 2011 Stanford Mass Spectrometry Users’ Meeting
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Healthy CarcinomaCirrhosis
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C57B6 female micemice
(6 mo.)
ildWild type (WT)
Sod1 Sod1 Sod1
HCV‐S
Sod1 Sod1 Sod1
HCV‐FL
Sod1 Sod1 Sod1Sod1
+/+
Sod1
‐/+
Sod1
‐/‐
Sod1
+/+
Sod1
‐/+
Sod1
‐/‐
Sod1
+/+
Sod1
‐/+
Sod1
‐/‐
Core E1 E2 p7 HCV-S
Core E1 E2 p7 NS3 NS4A NS4B NS5A NS5B HCV-FL
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Treatments:
Control vs Alcohol Biochemical ProteomicsControl vs. Alcohol diets
analyses Proteomics
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Treatments:
Control vs.
Biochemical analyses:
Liver enzyme activity; ProteomicsAlcohol
diet
Liver enzyme activity;
Histopathology; etc…
Proteomics
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Proteomics:
Treatments:
Control vs Alcohol
Biochemical analyses:
Liver enzyme activity;
Detecting protein modification via
bisANS or disulfide Control vs. Alcohol
diet
Liver enzyme activity;
HistopathologybisANS or disulfide
labeling;
2D electrophoresis; Mass Spectrometry
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15001400 -Control14001300
1200110010001000
900800
700600600500
400300200200100
0+/+ -/+ -/- +/+ -/+ -/- +/+ -/+ -/-
WT HCV-S HCV-FL
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15001400 - Control14001300
120011001000
- Alcohol
1000
900800
700600600500
400300200200100
0+/+ -/+ -/- +/+ -/+ -/- +/+ -/+ -/-
WT HCV-S HCV-FL
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Control
Sod1 +/+ Sod1 -/+ Sod1 -/-Normal liver
Alcohol
Sod1 +/+ Sod1 -/+ Sod1 -/-Fatty liver
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1 BisANS for protein conformational changes1. BisANS for protein conformational changes
1. 6‐IAF for protein modification
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2-D gel electrophoresiAdd bisANS UV irradiation
sAdd bisANS U ad a o
& cross linkProtein extraction
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SH
-SH
-S
ReducingAdd IAAProtein extraction
Reducing agent
Add IAA
-SH
-SH
2-D gel electrophoresi
sAdd 6-IAF
s
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Total proteinvia
Coomassie blue
Hydrophobicityvia
bisANS
Disulfide bondvia
6-IAF
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LC/MSD Trap XCT Ultra Agilent TechnologiesLC/MSD Trap XCT Ultra, Agilent Technologies
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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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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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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
![Page 26: For do not reuse or reproduce - Stanford University...For personal use only. Please do not reuse or reproduce without the author’s permission. 2 Presented at the 2011 Stanford Mass](https://reader035.vdocuments.us/reader035/viewer/2022062508/60a8876b64ba90168b350bfe/html5/thumbnails/26.jpg)
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
![Page 27: For do not reuse or reproduce - Stanford University...For personal use only. Please do not reuse or reproduce without the author’s permission. 2 Presented at the 2011 Stanford Mass](https://reader035.vdocuments.us/reader035/viewer/2022062508/60a8876b64ba90168b350bfe/html5/thumbnails/27.jpg)
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
![Page 28: For do not reuse or reproduce - Stanford University...For personal use only. Please do not reuse or reproduce without the author’s permission. 2 Presented at the 2011 Stanford Mass](https://reader035.vdocuments.us/reader035/viewer/2022062508/60a8876b64ba90168b350bfe/html5/thumbnails/28.jpg)
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
![Page 29: For do not reuse or reproduce - Stanford University...For personal use only. Please do not reuse or reproduce without the author’s permission. 2 Presented at the 2011 Stanford Mass](https://reader035.vdocuments.us/reader035/viewer/2022062508/60a8876b64ba90168b350bfe/html5/thumbnails/29.jpg)
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