The cDNA‐deduced primary structure of human sex hormone‐binding globulin and location of its steroid‐binding domain

FEBS Letters

Goping, I.S.²  Smith, C.L.²  Harley, M.J.²  Underhill, D.A.²  Cheng, C.Y.¹  Musto, N.A.¹  Bardin, C.W.¹  Hammond, G.L.² 

[1] The Population Council, 1230 York Avenue, New York, NY 10021, USA;Department of Obstetrics and Gynecology, University of Western Ontario, 375 South Street, London, Ontario N6A 4G5, Canada

关键词: cDNA cloning;    Amino acid sequence;    Sex hormone-binding globulin;    Steroid-binding domain;    SHBG;    sex hormone-binding globulin;    also known as testosterone-estradiol-binding globulin (TeBG) and sex-steroid-binding protein (SBP);    ABP;    androgen-binding protein;    Δ6-testosterone;    17β-hydroxyandrosta-4;    6-dien-3-one;    SSPE;    150 mM NaCl/10 mM NaH2PO4H2O/1 mM EDTA (pH 7.4);    SSC;    150 mM NaCl/15 mM sodium acetate (pH 7.4);    Denhardt's solution;    0.02% Ficoll/0.02% polyvinylpyrrolidone/0.02% bovine serum albumin (Pentax;    fraction V) in H2O (w/v);   

DOI  :  10.1016/0014-5793(87)80121-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have sequenced a cDNA for sex hormone-binding globulin (SHBG) isolated from a phage λgt 11 human liver cDNA library. The library was screened with a radiolabeled rat androgen-binding protein (ABP) cDNA, and the abundance of SHBG cDNAs was 1 in 750 000 plaques examined. The largest human SHBG cDNA (1194 base-pairs) contained a reading frame for 381 amino acids. This comprised 8 amino acids of a signal peptide followed by 373 residues starting with the known NH₂-terminal sequence of human SHBG, and ending with a termination codon. The predicted polypeptide M _r of SHBG is 40 509, and sites of attachment of one O-linked (residue 7) and two N-linked oligosaccharide (residues 351 and 367) chains were identified. Purified SHBG was photoaffinity-labeled with Δ⁶-[³H]testosterone and cleaved with trypsin. The labeled tryptic fragment was isolated by reverse-phase HPLC, and its NH₂-terminal sequence was determined. The results suggest that a portion of the steroid-binding domain of SHBG is located between residue 296 and the 35 predominantly hydrophilic residues at the C-terminus of the protein.

【 授权许可】

Unknown