Physicochemical characteristics of human sex hormone binding globulin: evidence for two identical subunits.
We have developed a rapid protocol for the purification of human sex hormone binding globulin (SHBG) which allows the protein to be purified from pregnancy serum within 48 h. This minimizes any possible degradation of the protein by serum proteases, and has enabled us to re-examine some important and controversial aspects of its structural composition. Our physicochemical data are consistent with the hypothesis that SHBG is a dimeric glycoprotein composed of 2 protomers that exhibit size heterogeneity (approximately 50 and approximately 52 K daltons). The dimeric SHBG molecule appears to contain only approximately 8% carbohydrate, and sequence information indicates that an N-linked oligosaccharide chain may be attached to residue 7 (asparagine) from the NH2-terminal amino acid (leucine). When compared with earlier reports, differences in the relative amounts of heavy (approximately 52 K) and light (approximately 50 K) protomers, and the microheterogeneity of NH2-terminal amino acids, have led us to conclude that they may be caused by proteolytic degradation in vivo as well as during the storage of blood samples prior to protein purification. However, the NH2-terminal amino acid sequence data indicate that the primary structures of the heavy protomers, which evidently interact to form the majority of SHBG dimer in serum, are similar and may even be identical. Evidence to support this is provided by the observation that a monoclonal antibody, which recognises a configurational epitope, interacts with two epitopes per native dimeric form of human SHBG.
G L
GL
Hammond
P A
PA
Robinson
H
H
Sugino
D N
DN
Ward
J
J
Finne
D000596:Amino Acids / Q000032:analysis; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002241:Carbohydrates / Q000032:analysis; D005260:Female; D006801:Humans; D046911:Macromolecular Substances; D008970:Molecular Weight; D011247:Pregnancy; D011263:Pregnancy Trimester, Third; D012738:Sex Hormone-Binding Globulin / Q000097:blood* / Q000302:isolation & purification
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002241:Carbohydrates; D046911:Macromolecular Substances; D012738:Sex Hormone-Binding Globulin