title: Resolution of the human sex hormone-binding globulin dimer interface and evidence for two steroid-binding sites per homodimer.
abstract: Human sex hormone-binding globulin (SHBG) transports sex steroids in the blood. It functions as a homodimer, but there is little information about the topography of its dimerization domain, and its steroid binding stoichiometry is controversial. The prevailing assumption is that each homodimeric SHBG molecule contains a single steroid-binding site at the dimer interface. However, crystallographic analysis of the amino-terminal laminin G-like domain of human SHBG has shown that the dimerization and steroid-binding sites are distinct and that both monomers within a homodimeric complex are capable of binding steroid. To validate our crystallographic model of the SHBG homodimer, we have used site-directed mutagenesis to create SHBG variants in which single amino acid substitutions (V89E and L122E) were introduced to produce steric clashes at critical positions within the proposed dimerization domain. The resulting dimerization-deficient SHBG variants contain a steroid-binding site with an affinity and specificity indistinguishable from wild-type SHBG. Moreover, when equalized in terms of their monomeric subunit content, dimerization-deficient and wild-type SHBGs have essentially identical steroid binding capacities. These data indicate that both subunits of the SHBG homodimer bind steroid and that measurements of the molar concentration of SHBG homodimer in serum samples have been overestimated by 2-fold.
authors.affiliation: Department of Obstetrics and Gynecology, The University of Western Ontario, London, Ontario N6A 4L6, Canada.
authors.forename: G V
authors.initials: GV
authors.lastname: Avvakumov
authors.forename: I
authors.initials: I
authors.lastname: Grishkovskaya
authors.forename: Y A
authors.initials: YA
authors.lastname: Muller
authors.forename: G L
authors.initials: GL
authors.lastname: Hammond
mesh_terms: D001665:Binding Sites; D019281:Dimerization; D006801:Humans; D012738:Sex Hormone-Binding Globulin / Q000032:analysis / Q000737:chemistry*; D013256:Steroids / Q000378:metabolism*
chemical_list: D012738:Sex Hormone-Binding Globulin; D013256:Steroids