The 56 kDa protein of human genital skin fibroblasts is identical to that radiolabelled by [3H]dihydrotestosterone 17 beta-bromoacetate.
Analysis of soluble proteins from human genital skin fibroblasts by two-dimensional polyacrylamide gel electrophoresis reveals an abundant protein doublet of mol. wt 56,000 with isoelectric points (pI) of 6.7 and 6.5. This protein is absent in non-genital skin fibroblasts as well as in genital skin fibroblasts of most patients with complete forms of androgen insensitivity. The protein specifically binds androgen. A protein of similar estimated molecular weight (58,000) from human genital skin fibroblasts has recently been found to be covalently radiolabelled by the affinity ligand dihydrotestosterone 17 beta-bromoacetate (DHT-BA). In the present study these proteins have been found to be indistinguishable on one- and two-dimensional gel electrophoresis. Antibodies raised against the 56 kDa pI 6.7/6.5 protein also recognized the protein covalently radiolabelled by DHT-BA. A third protein of estimated mol. wt 59,000 has been found to be associated with several steroid hormone receptor complexes but has no known ligand binding activity. This protein was found to be clearly separable from the 56/58 kDa protein on two-dimensional gel electrophoresis as it has a more acidic pI of approximately 5.4. Furthermore, antibodies against the 59 kDa protein do not recognize the 56 kDa species, and vice versa.
Belsham
D D
DD
Department of Biochemistry, University of Manitoba, Winnipeg, Canada.
Rosenmann
E
E
Pereira
F A
FA
Williams
S G
SG
Turney
M K
MK
Kovacs
W J
WJ
Faber
L E
LE
Wrogemann
K
K
D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D013196:Dihydrotestosterone / Q000031:analogs & derivatives*; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts / Q000032:analysis; D005835:Genitalia / Q000032:analysis*; D006801:Humans; D015151:Immunoblotting; D007120:Immunochemistry; D008970:Molecular Weight; D010777:Photochemistry; D010779:Photofluorography; D011506:Proteins / Q000032:analysis*; D011869:Radioligand Assay; D011944:Receptors, Androgen / Q000032:analysis*; D012867:Skin / Q000032:analysis
D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D011506:Proteins; D011944:Receptors, Androgen; D013196:Dihydrotestosterone; C023562:dihydrotestosterone 17-bromoacetate