Product of side-chain cleavage of cholesterol, isocaproaldehyde, is an endogenous specific substrate of mouse vas deferens protein, an aldose reductase-like protein in adrenocortical cells.
Mouse vas deferens protein (MVDP) is an aldose reductase-like protein that is highly expressed in the vas deferens and adrenal glands and whose physiological functions were unknown. We hereby describe the enzymatic characteristics of MVDP and its role in murine adrenocortical Y1 cells. The murine aldose reductase (AR) and MVDP cDNAs were expressed in bacteria to obtain recombinant proteins and to compare their enzymatic activities. Recombinant MVDP was functional and displayed kinetic properties distinct from those of murine AR toward various substrates, a preference for NADH, and insensitivity to AR inhibitors. For MVDP, isocaproaldehyde, a product of side-chain cleavage of cholesterol generated during steroidogenesis, is the best natural substrate identified so far. In Y1 cells, we found that NADH-linked isocaproaldehyde reductase (ICR) activity was much higher than NADPH-linked ICR activity and was not abolished by AR inhibitors. We demonstrate that in Y1 cells, forskolin-induced MVDP expression enhanced NADH-linked ICR activity by 5-6-fold, whereas no variation in ICR-linked NADPH activity was observed in the same experiment. In cells stably transfected with MVDP antisense cDNA, NADH-linked ICR activity was abolished even in the presence of forskolin, and the isocaproaldehyde toxicity was increased compared with that of intact Y1 cells, as measured by isocaproaldehyde LD(50). In Y1 cells transfected with MVDP antisense cDNA, forskolin-induced toxicity was abolished by aminoglutethimide. These results indicate that in adrenocortical cells, MVDP is responsible for detoxifying isocaproaldehyde generated by steroidogenesis.
Lefrançois-Martinez
A M
AM
UMR CNRS 6547, Université Blaise Pascal, Les Cézeaux, 63177 Aubière, Cedex, France.
Tournaire
C
C
Martinez
A
A
Berger
M
M
Daoudal
S
S
Tritsch
D
D
Veyssière
G
G
Jean
C
C
D000302:Adrenal Cortex / Q000201:enzymology*; D000449:Aldehyde Reductase / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism; D000616:Aminoglutethimide / Q000494:pharmacology; D000818:Animals; D002208:Caproates / Q000378:metabolism* / Q000494:pharmacology; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli; D006801:Humans; D008297:Male; D051379:Mice; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D016372:RNA, Antisense / Q000235:genetics / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014162:Transfection; D014649:Vas Deferens / Q000201:enzymology*
D000447:Aldehydes; D002208:Caproates; D004791:Enzyme Inhibitors; D011506:Proteins; D016372:RNA, Antisense; D011994:Recombinant Proteins; D000616:Aminoglutethimide; D009243:NAD; C018575:isocaproaldehyde; D005576:Colforsin; D009249:NADP; D002784:Cholesterol; C066498:Akr1b7 protein, mouse; D000449:Aldehyde Reductase; C027576:4-hydroxy-2-nonenal