Purification and characterization of the sex hormone-binding globulin in serum from Djungarian hamsters

A high-affinity sex hormone-binding globulin (SHBG) was purified from the serum of prepubertal Djungarian hamsters (Phodopus sungorus). A purification of more than 2000-fold with an overall yield of 23% was achieved without the use of androgen affinity chromatography. Two predominant variants (51 and 55 kDa) were resolved by denaturing polyacrylamide gel electrophoresis. Both variants participated in the binding of dihydrotestosterone (DHT) and had identical amino-terminal sequences. The sequences obtained for Djungarian hamster SHBG (dhSHBG) showed a high degree of identity with those of other mammals. The affinity of purified dhSHBG for DHT (2.5 x 10(9) M(-1) was similar to that measured in unfractionated serum. This protein was isolated as a dimer with a single calcium-dependent steroid-binding site and a major pI of 4.7. The described purification procedure yielded active dhSHBG from small volumes of prepubertal serum. These studies also provide the first direct structural evidence that a SHBG-like protein, not of testicular origin, is expressed by a rodent during prepubertal development.