Molecular cloning and amino acid sequence of the porcine 17 beta-estradiol dehydrogenase.
We describe the cloning and sequencing of porcine 17 beta-estradiol dehydrogenase. The enzyme performs oxidation 360-fold more efficiently than reduction, both measured under optimal conditions. It is localized in specialized vesicles of epithelial cells. The cDNA clones were isolated from a lambda UNI ZAP XR library of porcine kidney and polymerase-chain-reaction-amplified from templates of uterus epithelium. In both tissues, the same enzyme is coded by a transcript of 2.9 kb. It contains a 69-b 5'-noncoding region, an open reading frame of 2211 b and a 3'-noncoding region of 624 b. The open reading frame of 737 amino acids with a predicted molecular mass 79,973 Da was confirmed by amino acid sequencing of peptides. The 80-kDa translation product is processed to the N-terminal 32-kDa enzyme, part of which is then covalently linked to actin. The estradiol dehydrogenase/actin complex and the 80-kDa translation product comigrate in SDS/PAGE.
Leenders
F
F
Max-Planck-Institut für experimentelle Endokrinologie, Hannover, Germany.
Adamski
J
J
Husen
B
B
Thole
H H
HH
Jungblut
P W
PW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004960:Estradiol Dehydrogenases / Q000737:chemistry / Q000235:genetics*; D005260:Female; D007668:Kidney / Q000201:enzymology; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D013552:Swine; D014599:Uterus / Q000201:enzymology
D018076:DNA, Complementary; D004960:Estradiol Dehydrogenases