Rabbit sex hormone-binding globulin: expression in the liver and testis during postnatal development and structural characterization by truncated proteins.
Although sex hormone binding globulin (SHBG) is found in the blood plasma of adult humans and rabbits and the gene is expressed in their livers, it is not detected in the plasma of adult rodents nor is it expressed in adult rodent livers. Thus the rabbit represents a good model to study the metabolism and function of SHBG in the blood. We have used a cloned rabbit SHBG cDNA to detect mRNA expression in rabbits during the postnatal period, and to construct truncated SHBG proteins for structure/function analysis. The SHBG mRNA appeared in the testis as early as 3 days after birth. The level increased gradually in abundance throughout postnatal development, and attained a maximum at 12 weeks of age when the gonads were fully matured. In contrast, SHBG mRNA in the livers of male and female animals increased to a maximum by 4 weeks of age, and were maintained at this level until 12 weeks before subsiding to the initial levels. The increase and decrease in SHBG mRNA levels in the liver were accompanied by similar changes in serum SHBG. This suggests that SHBG in the blood circulation comes from the liver and this might also provide a source of SHBG for the male reproductive tract before formation of the blood-testis barrier. To elucidate the minimal sequence of rabbit SHBG responsible for steroid-binding, a panel of 13 truncated SHBG proteins was constructed, expressed in Escherichia coli, and biochemically purified for study. It was shown that the complete protein sequence of rabbit SHBG was important for maintaining a stable steroid-protein complex. Unlike human SHBG for which a truncated protein of the first 206 residues of the 373 amino acid protein can still bind steroid, removal of 43 or more residues from the C-terminus of rabbit SHBG completely abolished steroid-binding.
Wong
A S
AS
Department of Zoology, The University of Hong Kong, Pokfulam Road, Hong Kong, China.
Lui
W Y
WY
Hui
I T
IT
Lee
W M
WM
D000367:Age Factors; D000818:Animals; D001665:Binding Sites; D015152:Blotting, Northern; D018076:DNA, Complementary / Q000737:chemistry; D013196:Dihydrotestosterone / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005786:Gene Expression Regulation; D005982:Glutathione Transferase / Q000235:genetics; D007700:Kinetics; D008099:Liver / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*; D008297:Male; D023421:Models, Animal; D009154:Mutation; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012738:Sex Hormone-Binding Globulin / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D013737:Testis / Q000254:growth & development / Q000378:metabolism / Q000502:physiology*
D018076:DNA, Complementary; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012738:Sex Hormone-Binding Globulin; D013196:Dihydrotestosterone; D005982:Glutathione Transferase