Book chapter
DETECTION OF PROTEIN THIOLS IN MITOCHONDRIAL OXIDATIVE PHOSPHORYLATION COMPLEXES AND ASSOCIATED PROTEINS
Methods in Enzymology, pp.83-108
Methods in Enzymology, Elsevier
01/01/2010
DOI: 10.1016/S0076-6879(10)74006-4
PMID: 20609906
Abstract
The ability to detect and identify mitochondrial proteins that are sensitive to oxidative modification and inactivation by reactive species is important in understanding the molecular mechanisms responsible for mitochondrial dysfunction and tissue injury. In particular, cysteine residues play critical roles in maintaining the functional and structural integrity of numerous proteins in the mitochondrion and throughout the cell. To define changes in mitochondrial protein thiol status, proteomic approaches have been developed in which unmodified, reduced thiols (i.e., R-SH or thiolate species R-S(-)) are tagged with thiol-labeling reagents that can be visualized following gel electrophoresis and immunoblotting techniques. Herein, we describe the use of one thiol-labeling approach in combination with blue native gel electrophoresis (BN-PAGE) to detect reactive thiol groups within mitochondrial proteins including those of the oxidative phosphorylation (OxPhos) system. Labeling or "tagging" of protein thiol groups in combination with various gel electrophoresis and proteomics techniques is a valuable way to measure alterations in cellular or organelle thiol proteomes in response to drug treatment, disease state, or metabolic/oxidative stress.
Details
- Title: Subtitle
- DETECTION OF PROTEIN THIOLS IN MITOCHONDRIAL OXIDATIVE PHOSPHORYLATION COMPLEXES AND ASSOCIATED PROTEINS
- Creators
- Kelly K. Andringa - University of Alabama at BirminghamShannon M. Bailey - University of Alabama at Birmingham
- Contributors
- E Cadenas (Editor)L Packer (Editor)
- Resource Type
- Book chapter
- Publication Details
- Methods in Enzymology, pp.83-108
- Publisher
- Elsevier; SAN DIEGO
- Series
- Methods in Enzymology
- DOI
- 10.1016/S0076-6879(10)74006-4
- PMID
- 20609906
- eISSN
- 1557-7988
- ISSN
- 0076-6879
- Number of pages
- 26
- Grant note
- R21 DK073775; P30 DK74038; DK73775 / NIDDK NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Diabetes & Digestive & Kidney Diseases (NIDDK) S10 RR11329; S10 RR13795 / NCRR NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Center for Research Resources (NCRR) P30 AR050948 / NIAMS NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Arthritis & Musculoskeletal & Skin Diseases (NIAMS) AA15172; R01 AA015172 / NIAAA NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute on Alcohol Abuse & Alcoholism (NIAAA) P50AT000477 / NATIONAL CENTER FOR COMPLEMENTARY &ALTERNATIVE MEDICINE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Center for Complementary & Alternative Medicine S10RR013795 / NATIONAL CENTER FOR RESEARCH RESOURCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Center for Research Resources (NCRR) P30AR050948 / NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Arthritis & Musculoskeletal & Skin Diseases (NIAMS) P30DK074038 / NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Diabetes & Digestive & Kidney Diseases (NIDDK) P50 AT00477 / NCCIH NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Center for Complementary & Alternative Medicine R01AA015172 / NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute on Alcohol Abuse & Alcoholism (NIAAA) U54 CA100949; P30 CA13148 / NCI NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) P30CA013148 / NATIONAL CANCER INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI)
- Language
- English
- Date published
- 01/01/2010
- Academic Unit
- Orthopedics and Rehabilitation
- Record Identifier
- 9984548856202771
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