An androgenic affinity ligand covalently binds to cytosolic aldehyde dehydrogenase from human genital skin fibroblasts.
A 56 kDa protein expressed in human genital skin fibroblasts was first identified by independent laboratories on the basis of its specific expression in androgen target cells and its ability to covalently bind androgenic affinity ligands. Recently, immunoscreening of a cDNA library with antisera directed against this protein resulted in the isolation of a partial cDNA clone identical to human cytosolic aldehyde dehydrogenase (ALDH1). We report here the preparation of a full-length cDNA encoding ALDH1 from human genital fibroblasts. Translation of the encoded protein in a cell-free system yields a 56 kDa product that can be covalently radiolabeled with [3H]dihydrotestosterone 17 beta-bromoacetate (DHT-BA). Expression of the full-length clone in mammalian cells also results in expression of a 56 kDa DHT-BA binding protein. The covalent binding of DHT-BA by ALDH1 is an intrinsic property of the enzyme and is not dependent on androgen receptor expression.
McCammon
D K
DK
Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232.
Zhou
P
P
Turney
M K
MK
McPhaul
M J
MJ
Kovacs
W J
WJ
D000345:Affinity Labels; D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D000728:Androgens / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D016466:CHO Cells; D006224:Cricetinae; D004247:DNA / Q000737:chemistry / Q000235:genetics; D013196:Dihydrotestosterone / Q000031:analogs & derivatives / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D005835:Genitalia / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D014162:Transfection
D000345:Affinity Labels; D000728:Androgens; D013196:Dihydrotestosterone; C023562:dihydrotestosterone 17-bromoacetate; D004247:DNA; D000444:Aldehyde Dehydrogenase