Open AccessMolecular cloning and amino acid sequence of the porcine 17β‐estradiol dehydrogenase
Author(s) -
LEENDERS F.,
ADAMSKI J.,
HUSEN B.,
THOLE H. H.,
JUNGBLUT P. W.
Publication year - 1994
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1994.tb18860.x
Subject(s) - open reading frame , complementary dna , microbiology and biotechnology , amino acid , peptide sequence , biochemistry , biology , molecular cloning , dehydrogenase , cloning (programming) , enzyme , chemistry , gene , computer science , programming language
We describe the cloning and sequencing of porcine 17β‐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 λUNI 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 79973 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.