CDNA CLONING AND CHARACTERIZATION OF A NEW HUMAN MICROSOMAL NAD(-DEPENDENT DEHYDROGENASE THAT OXIDIZES ALL-TRANS-RETINOL AND 3-ALPHA-HYDROXYSTEROIDS())
Wh. Gough et al., CDNA CLONING AND CHARACTERIZATION OF A NEW HUMAN MICROSOMAL NAD(-DEPENDENT DEHYDROGENASE THAT OXIDIZES ALL-TRANS-RETINOL AND 3-ALPHA-HYDROXYSTEROIDS()), The Journal of biological chemistry, 273(31), 1998, pp. 19778-19785
We report the cDNA sequence and catalytic properties of a new member o
f the short chain dehydrogenase/reductase superfamily, The 1134-base p
air cDNA isolated from the human liver cDNA library encodes a 317-amin
o acid protein, retinol dehydrogenase 4 (RoDH-4), which exhibits the s
trongest similarity with rat all-trans-retinol dehydrogenases RoDH-1,
RoDH-2, and RoDH-3, and mouse cis-retinol/androgen dehydrogenase (less
than or equal to 73% identity). The mRNA for RoDH-4 is abundant in ad
ult liver, where it is translated into RoDH-4 protein, which is associ
ated with microsomal membranes, as evidenced by Western blot analysis,
Significant amounts of RoDH-4 message are detected in fetal liver and
lung. Recombinant RoDH-4, expressed in microsomes of Sf9 insect cells
using BacoluGold Baculovirus system, oxidizes all-trans retinol and 1
3-cis-retinol to corresponding aldehydes and oxidizes the 3 alpha-hydr
oxysteroids androstane-diol and androsterone to dihydrotestosterone an
d androstanedione, respectively. NAD(+) and NADH are the preferred cof
actors, with apparent K-m values 250-1500 times lower than those for N
ADP(+) and NADPH. AU-trans-retinol and 13-cis-retinol inhibit RoDH-4 c
atalyzed oxidation of androsterone with apparent K-i values of 5.8 and
3.5 mu M, respectively. All-trans-retinol bound to cellular retinol-b
inding protein (type I) exhibits a similar K-i value of 3.6 mu M, Unli
ganded cellular retinol-binding protein has no effect on RoDH activity
. Citral and acyclic isoprenoids also act as inhibitors of RoDH-4 acti
vity. Ethanol is not inhibitory. Thus, we have identified and characte
rized a sterol/retinol-oxidizing short chain dehydrogenase/reductase t
hat prefers NAD(+) and recognizes all-trans-retinol as substrate, RoDH
-4 can potentially contribute to the biosynthesis of two powerful modu
lators of gene expression: retinoic acid from retinol and dihydrotesto
sterone from 3 alpha-androstane-diol.