Coenzyme isomerization is integral to catalysis in aldehyde dehydrogenase

Samantha J. Perez-Miller, Thomas Hurley

Research output: Contribution to journalArticle

122 Citations (Scopus)

Abstract

Crystal structures of many enzymes in the aldehyde dehydrogenase superfamily determined in the presence of bound NAD(P)+ have exhibited conformational flexibility for the nicotinamide half of the cofactor. This has been hypothesized to be important in catalysis because one conformation would block the second half of the reaction, but no firm evidence has been put forth which shows whether the oxidized and reduced cofactors preferentially occupy the two observed conformations. We present here two structures of the wild type and two structures of a Cys302Ser mutant of human mitochondrial aldehyde dehydrogenase in binary complexes with NAD+ and NADH. These structures, including the Cys302Ser mutant in complex with NAD+ at 1.4 Å resolution and the wild-type enzyme in complex with NADH at 1.9 Å resolution, provide strong evidence that bound NAD+ prefers an extended conformation ideal for hydride transfer and bound NADH prefers a contracted conformation ideal for acyl - enzyme hydrolysis. Unique interactions between the cofactor and the Rossmann fold make isomerization possible while allowing the remainder of the active site complex to remain intact. In addition, these structures clarify the role of magnesium in activating the human class 2 enzyme. Our data suggest that the presence of magnesium may lead to selection of particular conformations and speed isomerization of the reduced cofactor following hydride transfer.

Original languageEnglish
Pages (from-to)7100-7109
Number of pages10
JournalBiochemistry
Volume42
Issue number23
DOIs
StatePublished - Jun 17 2003

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Aldehyde Dehydrogenase
Coenzymes
Isomerization
Catalysis
NAD
Conformations
Enzymes
Hydrides
Magnesium
Niacinamide
Hydrolysis
Catalytic Domain
Crystal structure

ASJC Scopus subject areas

  • Biochemistry

Cite this

Coenzyme isomerization is integral to catalysis in aldehyde dehydrogenase. / Perez-Miller, Samantha J.; Hurley, Thomas.

In: Biochemistry, Vol. 42, No. 23, 17.06.2003, p. 7100-7109.

Research output: Contribution to journalArticle

Perez-Miller, Samantha J. ; Hurley, Thomas. / Coenzyme isomerization is integral to catalysis in aldehyde dehydrogenase. In: Biochemistry. 2003 ; Vol. 42, No. 23. pp. 7100-7109.
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