Identification and Quantification of Protein Carbonylation by Mass Spectrometry

Accumulated evidence indicates oxidative stress plays important roles in disease and aging. Under oxidative stress, lipid peroxidation (LPO) leads to reactive carbonyl species (RCS) that can modify a wide range of biomolecules including protein, DNA and carbohydrate. In this dissertation, we investigate the modification of two model proteins, human serum albumin (HSA) and aconitase (ACO), by the LPO-relevant a, b-unsaturated aldehydes, acrolein (ACR) and 4-hydroxy-2-nonenal (HNE). The investigation is focused on the characterization and quantification ACR and HNE addition to the model proteins. A correlation between HNE modification and ACO activity is also determined. These results provide insights into the impact of oxidative stress at the molecular level and are relevant to aging and disease states. We finally investigate protein carbonylation in ischemic mouse heart mitochondria, and develop a quantitative method for detecting carbonylated protein in this system. The research is based on liquid chromatography/mass spectrometry (LC/MS), Western Blots, and enzymatic assay.

Name thesis:degree_name

Doctor of Philosophy

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Chemistry

Year dc:date.available

2012

Author dc:creator

• Liu, Qingyuan

Contributors dc:contributor

• Scott Gronert

• protein carbonylation
• mass spectrometry
• Chemistry
• Physical Sciences and Mathematics

Statement dc:rights

• © The Author

Identifier

https://scholarscompass.vcu.edu/etd/321

OAI identifier oai:identifier

oai:scholarscompass.vcu.edu:etd-1320