PMID- 15950210
OWN - NLM
STAT- MEDLINE
DCOM- 20050823
LR  - 20161124
IS  - 0009-2797 (Print)
IS  - 0009-2797 (Linking)
VI  - 155
IP  - 1-2
DP  - 2005 Jun 30
TI  - The interactions of 9,10-phenanthrenequinone with glyceraldehyde-3-phosphate 
      dehydrogenase (GAPDH), a potential site for toxic actions.
PG  - 97-110
AB  - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) catalyzes the oxidative 
      phosphorylation of glyceraldehyde 3-phosphate to 1,3-diphosphoglycerate, one of 
      the precursors for glycolytic ATP biosynthesis. The enzyme contains an active 
      site cysteine thiolate, which is critical for its catalytic function. As part of 
      a continuing study of the interactions of quinones with biological systems, we 
      have examined the susceptibility of GAPDH to inactivation by 
      9,10-phenanthrenequinone (9,10-PQ). In a previous study of quinone toxicity, this 
      quinone, whose actions have been exclusively attributed to reactive oxygen 
      species (ROS) generation, caused a reduction in the glycolytic activity of GAPDH 
      under aerobic and anaerobic conditions, indicating indirect and possible direct 
      actions on this enzyme. In this study, the effects of 9,10-PQ on GAPDH were 
      examined in detail under aerobic and anaerobic conditions so that the role of 
      oxygen could be distinguished from the direct effects of the quinone. The results 
      indicate that, in the presence of the reducing agent DTT, GAPDH inhibition by 
      9,10-PQ under aerobic conditions was mostly indirect and comparable to the direct 
      actions of exogenously-added H2O2 on this enzyme. GAPDH was also inhibited by 
      9,10-PQ anaerobically, but in a somewhat more complex manner. This quinone, which 
      is not considered an electrophile, inhibited GAPDH in a time-dependent manner, 
      consistent with irreversible modification and comparable to the electrophilic 
      actions of 1,4-benzoquinone (1,4-BQ). Analysis of the anaerobic inactivation 
      kinetics for the two quinones revealed comparable inactivation rate constants 
      (k(inac)), but a much lower inhibitor binding constant (K(i)) for 1,4-BQ. 
      Protection and thiol titration studies suggest that these quinones bind to the 
      NAD+ binding site and modify the catalytic thiol from this site. Thus, 9,10-PQ 
      inhibits GAPDH by two distinct mechanisms: through ROS generation that results in 
      the oxidization of GAPDH thiols, and by an oxygen-independent mechanism that 
      results in the modification of GAPDH catalytic thiols.
FAU - Rodriguez, Chester E
AU  - Rodriguez CE
AD  - Department of Pharmacology, UCLA School of Medicine, Center for the Health 
      Sciences, Los Angeles, CA 90095-1735, USA.
FAU - Fukuto, Jon M
AU  - Fukuto JM
FAU - Taguchi, Keiko
AU  - Taguchi K
FAU - Froines, John
AU  - Froines J
FAU - Cho, Arthur K
AU  - Cho AK
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - Ireland
TA  - Chem Biol Interact
JT  - Chemico-biological interactions
JID - 0227276
RN  - 0 (Air Pollutants)
RN  - 0 (Benzoquinones)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Mutagens)
RN  - 0 (Phenanthrenes)
RN  - 3T006GV98U (quinone)
RN  - 42L7BZ8H74 (9,10-phenanthrenequinone)
RN  - BBX060AN9V (Hydrogen Peroxide)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
SB  - IM
MH  - Air Pollutants
MH  - Benzoquinones/toxicity
MH  - Dose-Response Relationship, Drug
MH  - Enzyme Inhibitors/*toxicity
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/*antagonists & inhibitors
MH  - Hydrogen Peroxide/toxicity
MH  - Mutagens/*toxicity
MH  - Oxidation-Reduction
MH  - Phenanthrenes/*toxicity
MH  - Saccharomyces cerevisiae/enzymology
EDAT- 2005/06/14 09:00
MHDA- 2005/08/24 09:00
CRDT- 2005/06/14 09:00
PHST- 2005/02/11 00:00 [received]
PHST- 2005/05/10 00:00 [revised]
PHST- 2005/05/10 00:00 [accepted]
PHST- 2005/06/14 09:00 [pubmed]
PHST- 2005/08/24 09:00 [medline]
PHST- 2005/06/14 09:00 [entrez]
AID - S0009-2797(05)00151-1 [pii]
AID - 10.1016/j.cbi.2005.05.002 [doi]
PST - ppublish
SO  - Chem Biol Interact. 2005 Jun 30;155(1-2):97-110. doi: 10.1016/j.cbi.2005.05.002.