PMID- 20609906
OWN - NLM
STAT- MEDLINE
DCOM- 20101029
LR  - 20211020
IS  - 1557-7988 (Electronic)
IS  - 0076-6879 (Linking)
VI  - 474
DP  - 2010
TI  - Detection of protein thiols in mitochondrial oxidative phosphorylation complexes 
      and associated proteins.
PG  - 83-108
LID - 10.1016/S0076-6879(10)74006-4 [doi]
AB  - 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.
CI  - Copyright (c) 2010 Elsevier Inc. All rights reserved.
FAU - Andringa, Kelly K
AU  - Andringa KK
AD  - Department of Environmental Health Sciences, Center for Free Radical Biology, 
      University of Alabama at Birmingham, Birmingham, Alabama, USA.
FAU - Bailey, Shannon M
AU  - Bailey SM
LA  - eng
GR  - P30 AR050948/AR/NIAMS NIH HHS/United States
GR  - DK73775/DK/NIDDK NIH HHS/United States
GR  - P30 DK74038/DK/NIDDK NIH HHS/United States
GR  - P50 AT00477/AT/NCCIH NIH HHS/United States
GR  - R01 AA015172/AA/NIAAA NIH HHS/United States
GR  - S10 RR13795/RR/NCRR NIH HHS/United States
GR  - R21 DK073775/DK/NIDDK NIH HHS/United States
GR  - P30 CA13148/CA/NCI NIH HHS/United States
GR  - U54 CA100949/CA/NCI NIH HHS/United States
GR  - AA15172/AA/NIAAA NIH HHS/United States
GR  - S10 RR11329/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20100620
PL  - United States
TA  - Methods Enzymol
JT  - Methods in enzymology
JID - 0212271
RN  - 0 (Mitochondrial Proteins)
RN  - 0 (Sulfhydryl Compounds)
SB  - IM
MH  - Animals
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Humans
MH  - Immunoblotting
MH  - Mass Spectrometry
MH  - Mitochondria, Liver/enzymology
MH  - Mitochondrial Proteins/*chemistry
MH  - Oxidative Phosphorylation
MH  - Proteomics
MH  - *Staining and Labeling/methods
MH  - Sulfhydryl Compounds/*analysis/chemistry
EDAT- 2010/07/09 06:00
MHDA- 2010/10/30 06:00
CRDT- 2010/07/09 06:00
PHST- 2010/07/09 06:00 [entrez]
PHST- 2010/07/09 06:00 [pubmed]
PHST- 2010/10/30 06:00 [medline]
AID - S0076-6879(10)74006-4 [pii]
AID - 10.1016/S0076-6879(10)74006-4 [doi]
PST - ppublish
SO  - Methods Enzymol. 2010;474:83-108. doi: 10.1016/S0076-6879(10)74006-4. Epub 2010 
      Jun 20.