PMID- 12069614
OWN - NLM
STAT- MEDLINE
DCOM- 20020712
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 41
IP  - 25
DP  - 2002 Jun 25
TI  - Identification of bovine heart cytochrome c oxidase subunits modified by the 
      lipid peroxidation product 4-hydroxy-2-nonenal.
PG  - 8212-20
AB  - Bovine heart cytochrome c oxidase (CcO) was inactivated by the lipid peroxidation 
      product 4-hydroxy-2-nonenal (HNE) in a time- and concentration-dependent manner 
      with pseudo-first-order kinetics. Cytochrome c oxidase electron transport 
      activity decreased by as much as 50% when the enzyme was incubated for 2 h at 
      room temperature with excess HNE (300-500 microM). HNE-modified CcO subunits were 
      identified by two mass spectrometric methods: electrospray ionization mass 
      spectrometry (ESI/MS) and matrix-assisted laser desorption/ionization 
      time-of-flight mass spectrometry (MALDI-TOF/MS). All of the experimentally 
      determined molecular masses were in excellent agreement with published sequence 
      values with an accuracy of approximately 1 part per 10000 mass units for subunits 
      smaller than 20 kDa and approximately 1 part per 1000 mass units for the three 
      subunits larger than 20 kDa. Both MS methods detected six CcO subunits with an 
      increased mass of 156 Da after reaction with HNE (subunits II, IV, Vb, VIIa, 
      VIIc, and VIII); this result indicates a single Michael-type reaction site on 
      either a lysine or histidine residue within each subunit. Reaction of HNE with 
      either subunit VIIc or subunit VIII (modified approximately 30% and 50-75%, 
      respectively) must be responsible for CcO inhibition. None of the other subunits 
      were modified more than 5% and could not account for the observed loss of 
      activity. Reaction of HNE with His-36 of subunit VIII is most consistent with the 
      approximately 50% inhibition of CcO: (1) subunit VIII is modified more than any 
      other subunit by HNE; (2) the time dependence of subunit VIII modification is 
      consistent with the percent inhibition of CcO; (3) His-36 was identified as the 
      HNE-modified amino acid residue within subunit VIII by tandem MS analysis.
FAU - Musatov, Andrej
AU  - Musatov A
AD  - Department of Biochemistry, The University of Texas Health Science Center, 7703 
      Floyd Curl Drive, San Antonio, TX 78229-3900, USA. musatov@uthscsa.edu
FAU - Carroll, Christopher A
AU  - Carroll CA
FAU - Liu, Yuan-Chao
AU  - Liu YC
FAU - Henderson, George I
AU  - Henderson GI
FAU - Weintraub, Susan T
AU  - Weintraub ST
FAU - Robinson, Neal C
AU  - Robinson NC
LA  - eng
GR  - AA 10114/AA/NIAAA NIH HHS/United States
GR  - GM S 24795/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Aldehydes)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Reactive Oxygen Species)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
RN  - K1CVM13F96 (4-hydroxy-2-nonenal)
SB  - IM
MH  - Aldehydes/*metabolism/pharmacology
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cattle
MH  - Chromatography, High Pressure Liquid
MH  - Electron Transport Complex IV/antagonists & inhibitors/*metabolism
MH  - Enzyme Inhibitors/*metabolism/pharmacology
MH  - *Lipid Peroxidation/drug effects
MH  - Molecular Sequence Data
MH  - Myocardium/*enzymology/metabolism
MH  - Oxidative Stress
MH  - Reactive Oxygen Species/metabolism
MH  - Spectrometry, Mass, Electrospray Ionization
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
EDAT- 2002/06/19 10:00
MHDA- 2002/07/13 10:01
CRDT- 2002/06/19 10:00
PHST- 2002/06/19 10:00 [pubmed]
PHST- 2002/07/13 10:01 [medline]
PHST- 2002/06/19 10:00 [entrez]
AID - bi025896u [pii]
AID - 10.1021/bi025896u [doi]
PST - ppublish
SO  - Biochemistry. 2002 Jun 25;41(25):8212-20. doi: 10.1021/bi025896u.