Purification of the sex steroid binding protein from human serum
- PMID: 164886
- DOI: 10.1021/bi00676a013
Purification of the sex steroid binding protein from human serum
Abstract
The sex steroid binding protein from human pregnancy serum was purified to homogeneity by affinity chromatography and preparative polyacrylamide gel electrophoresis. The selective adsorbants were prepared by coupling [3H]-5alpha-dihydrotestosterone 17beta-hemisuccinate to 3,3'-diaminodipropylamine-agarose, poly(Lys-DLAla)-agarose, and albumin-agarose. The most effective adsorbant purifying for the binding protein was 5alpha-dihydrotestosterone 17beta-hemisuccinyl-3,3'-diaminodipropylamine-agarose. A preparative procedure with 5alpha-dihydrotestosterone 17beta-hemisuccinyl-3,3'-diaminodipropylamine-agarose yielded active material which was further purified by preparative polyacrylamide electrophoresis at pH 9.5. Homogeneity was shown by analytical disc gel electrophoresis at three different pH units. A single radioactive band corresponding to the stained band was shown by incubating with [1,2-3H]-5alpha-dihydrotestosterone prior to electrophoresis. The radioactive peak corresponding to the pure sex steroid binding protein could not be detected when a 100-fold excess of 17beta-estradiol was present in the incubation prior to electrophoresis demonstrating the specific sex steroid binding properties of this protein. The migration of this peak was identical with that obtained when diluted serum was electrophoresed under the same conditions in the presence of [1,2-3H]-5alpha-dihydrotestosterone indicating that no significant changes in the molecular characteristics of the binding protein occurred during the purification procedure. The presence of carbohydrate in the pure protein was shown by the periodic acid-Schiff reagent procedure. Selective adsorbants containing 17beta-estradiol linked at the 3 position were ineffective in retaining sex steroid binding protein activity.
Similar articles
-
Characterization of the sex steroid binding protein of human pregnancy serum. Improvements in the purification procedure.Biochemistry. 1978 Apr 18;17(8):1409-15. doi: 10.1021/bi00601a008. Biochemistry. 1978. PMID: 565650
-
The use of deoxyribonucleic acid-cellulose chromatography and isoelectric focusing for the characterization and partial purification of steroid-receptor complexes.Biochem J. 1973 May;134(1):113-27. doi: 10.1042/bj1340113. Biochem J. 1973. PMID: 4353081 Free PMC article.
-
Purification and characterization of the sex steroid-binding protein of rabbit serum. Comparison with the human protein.J Biol Chem. 1978 Aug 10;253(15):5293-8. J Biol Chem. 1978. PMID: 566754
-
Characterization of the androgen receptor in the anterior pituitary of the rat.Endocrinology. 1975 Dec;97(6):1355-63. doi: 10.1210/endo-97-6-1355. Endocrinology. 1975. PMID: 173521
-
2-Iodoestradiol binds with high affinity to human sex hormone binding globulin (SHBG).J Steroid Biochem. 1990 Jun;36(1-2):75-81. doi: 10.1016/0022-4731(90)90115-9. J Steroid Biochem. 1990. PMID: 2362451
Cited by
-
Effect of sex and prior exposure to a cafeteria diet on the distribution of sex hormones between plasma and blood cells.PLoS One. 2012;7(3):e34381. doi: 10.1371/journal.pone.0034381. Epub 2012 Mar 27. PLoS One. 2012. PMID: 22479617 Free PMC article.
-
Sertoli cells of adult rats in vitro. III. Purification of androgen-binding protein from the culture medium.Experientia. 1982 Mar 15;38(3):406-7. doi: 10.1007/BF01949420. Experientia. 1982. PMID: 7200433
-
Purification of foetal steroid-binding protein from human serum by affinity chromatography on 5 alpha-androstane-3 beta,17 beta-diol 3-hemisuccinate-aminohexyl-Sepharose 6B.Biochem J. 1986 Nov 15;240(1):75-9. doi: 10.1042/bj2400075. Biochem J. 1986. PMID: 3827855 Free PMC article.
-
Steroid hormone receptors in the regulation of differentiation. A review.Am J Pathol. 1977 Mar;86(3):705-44. Am J Pathol. 1977. PMID: 190892 Free PMC article. Review. No abstract available.
-
Hormonal and immunological aspects of the phylogeny of sex steroid binding plasma protein.Proc Natl Acad Sci U S A. 1980 Aug;77(8):4578-82. doi: 10.1073/pnas.77.8.4578. Proc Natl Acad Sci U S A. 1980. PMID: 6933505 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Other Literature Sources