Cloning of a cDNA encoding an aldehyde dehydrogenase and its expression in Escherichia coli: Recognition of retinal as substrate

Xianshu Wang, Peter Penzes, Joseph L. Napoli*

*Corresponding author for this work

Research output: Contribution to journalArticle

148 Scopus citations

Abstract

The biosynthesis of the hormone retinoic acid from retinol (vitamin A) involves two sequential steps, catalyzed by retinol dehydrogenases and retinal dehydrogenases, respectively. This report describes the cloning of a cDNA encoding a heretofore unknown aldehyde dehydrogenase from a rat testis library and its expression in Escherichia coli. This enzyme has been designated retinal dehydrogenase, type II, RalDH(II). The deduced amine acid sequence of RalDH(II) had the highest identity with mammalian aldehyde dehydrogenases that feature low K(m) values (μM) for retinal: human ALDH1 (72.2%), rat retinal dehydrogenase, type I (71.5%), bovine retina (72.7%), and mouse AHD-2 (71.5%). RalDH(II) expressed in E. coli recognizes as substrates free retinal, with a K(m) of ~0.7 μM, and cellular retinol- binding protein-bound retinal, with a K(m) of ~0.2 μM. RalDH(II) also can utilize as substrate retinal generated in situ by microsomal retinol dehydrogenases, from the physiologically most abundant substrate: retinol bound to cellular retinol-binding protein. Rat testis expresses RalDH(II) mRNA most abundantly, followed by (relative to testis): lung (6.7%), brain (6.3%), heart (5.2%), liver (4.4%), and kidney (2.7%). RalDH(II) does not recognize citral, benzaldehyde, acetaldehyde, and propanal efficiently as substrates, but does metabolize octanal and decanal efficiently. These data support a function for RalDH(II) in the pathway of retinoic acid biogenesis.

Original languageEnglish (US)
Pages (from-to)16288-16293
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number27
DOIs
StatePublished - Jul 29 1996

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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