Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1976 Dec 1;159(3):651–659. doi: 10.1042/bj1590651

Purification and properties of bovine pituitary follitropin.

K W Cheng
PMCID: PMC1164165  PMID: 1008823

Abstract

A reproducible procedure was developed for the purification of follitropin from frozen bovine pituitary glands. The method involved precipitation with (NH4)2SO4 and acetone, followed by ion-exchange column chromatography on CM-cellulose and DEAE-cellulose and gel filtration on Sephadex G-100. A specific radioligand-receptor assay for follitropin was used to locate the activity in eluates after column chromatography and gel filtration. The potency of the highly purified bovine follitropin as measured by Steelman-Pohley bioassay was 164 times that of NIH-FSH-S1 standard preparation. They yield of bovine follitropin was 2.9 mg/kg of frozen pituitary glands. Electrophoretically, bovine follitropin was more acidic in nature and migrated further towards the anode than lutropin and thyrotropin. The elution volume of bovine follitropin by gel filtration on Sephadex G-100 was very similar to that of bovine lutropin. The amino acid composition of bovine follitropin was similar to that of sheep and human follitropin, being rich in lysine, aspartic acid, threonine, serine, glutamic acid and half-cystine.

Full text

PDF
652

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amir S. M., Parlow A. F., Sawyer C. H. Amino acid composition and molecular weight of human follicle stimulating hormone. Proc Soc Exp Biol Med. 1972 Apr;139(4):1120–1122. doi: 10.3181/00379727-139-36311. [DOI] [PubMed] [Google Scholar]
  2. Andrews P. Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochem J. 1964 May;91(2):222–233. doi: 10.1042/bj0910222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BATES R. W., CONDLIFFE P. G. Chromatography of thyroid-stimulating hormone on carboxymethylcellulose. J Biol Chem. 1956 Dec;223(2):843–852. [PubMed] [Google Scholar]
  4. Braselton W. E., Jr, McShan W. H. Purification and properties of follicle-stimulating and luteinizing hormones from horse pituitary glands. Arch Biochem Biophys. 1970 Jul;139(1):45–58. doi: 10.1016/0003-9861(70)90043-3. [DOI] [PubMed] [Google Scholar]
  5. CONDLIFFE P. G., BATES R. W., FRAPS R. M. Fractionation of bovine thyrotrophin and luteinizing hormone on cellulose ion exchange columns. Biochim Biophys Acta. 1959 Aug;34:430–438. doi: 10.1016/0006-3002(59)90295-1. [DOI] [PubMed] [Google Scholar]
  6. Cahill C. L., Shetlar M. R., Payne R. W., Endecott B., Li Y. T. Isolation and characterization of ovine follicle stimulating hormone. Biochim Biophys Acta. 1968 Jan 22;154(1):40–52. doi: 10.1016/0005-2795(68)90256-0. [DOI] [PubMed] [Google Scholar]
  7. Cheng K. W. A radioreceptor assay for follicle-stimulating hormone. J Clin Endocrinol Metab. 1975 Sep;41(3):581–589. doi: 10.1210/jcem-41-3-581. [DOI] [PubMed] [Google Scholar]
  8. Cheng K. W. Carboxymethylation of methionine residues in bovine pituitary luteinizing hormone and its subunits. Effects on the binding activity with receptor sites and interactions between subunits. Biochem J. 1976 Oct 1;159(1):71–77. doi: 10.1042/bj1590071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cheng K. W. Properties of follicle-stimulating-hormone receptor in cell membranes of bovine testis. Biochem J. 1975 Jul;149(1):123–132. doi: 10.1042/bj1490123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  11. ELLIS S. A scheme for the separation of pituitary proteins. J Biol Chem. 1958 Jul;233(1):63–68. [PubMed] [Google Scholar]
  12. ELLIS S. Studies on the serial extraction of pituitary proteins. Endocrinology. 1961 Sep;69:554–570. doi: 10.1210/endo-69-3-554. [DOI] [PubMed] [Google Scholar]
  13. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  14. Hartree A. S. Purification of human pituitary glycoprotein hormones. Methods Enzymol. 1975;37:380–389. doi: 10.1016/s0076-6879(75)37033-x. [DOI] [PubMed] [Google Scholar]
  15. Hermier C., Jutisz M. Obtention de l'hormone folliculo-stimulante de mouton de pureté biologique élevée par la technique du recyclage sur gel Séphadex. Biochim Biophys Acta. 1969 Mar;175(2):402–408. [PubMed] [Google Scholar]
  16. Papkoff H., Gospodarowicz D., Li C. H. Purification and properties of ovine follicle-stimulating hormone. Arch Biochem Biophys. 1967 May;120(2):434–439. doi: 10.1016/0003-9861(67)90261-5. [DOI] [PubMed] [Google Scholar]
  17. Papkoff H., Mahlmann L. J., Li C. H. Some chemical and physical properties of human pituitary follicle-stimulating hormone. Biochemistry. 1967 Dec;6(12):3976–3982. doi: 10.1021/bi00864a045. [DOI] [PubMed] [Google Scholar]
  18. Peckham W. D., Parlow A. F. Isolation from human pituitary glands of three discrete electrophoretic components with high luteinizing hormone activity. Endocrinology. 1969 Sep;85(3):618–622. doi: 10.1210/endo-85-3-618. [DOI] [PubMed] [Google Scholar]
  19. Pierce J. G., Liao T., Howard S. M., Shome B., Cornell J. S. Studies on the structure of thyrotropin: its relationship to luteinizing hormone. Recent Prog Horm Res. 1971;27:165–212. doi: 10.1016/b978-0-12-571127-2.50029-8. [DOI] [PubMed] [Google Scholar]
  20. ROOS P., GEMZELL C. A. THE ISOLATION OF HUMAN URINARY FOLLICLE-STIMULATING HORMONE. Biochim Biophys Acta. 1964 Oct 9;93:217–220. doi: 10.1016/0304-4165(64)90291-0. [DOI] [PubMed] [Google Scholar]
  21. Rathnam P., Saxena B. B. Primary amino acid sequence of follicle-stimulating hormone from human pituitary glands. I. alpha subunit. J Biol Chem. 1975 Sep 10;250(17):6735–6746. [PubMed] [Google Scholar]
  22. Reichert L. E., Jr Electrophoretic properties of pituitary gonadotropins as studied by electrofocusing. Endocrinology. 1971 Apr;88(4):1029–1044. doi: 10.1210/endo-88-4-1029. [DOI] [PubMed] [Google Scholar]
  23. Reichert L. E., Jr, Kathan R. H., Ryan R. J. Studies on the composition and properties of immunochemical grade human pituitary follicle stimulating hormone (FSH): comparison with luteinizing hormone (LH). Endocrinology. 1968 Jan;82(1):109–114. doi: 10.1210/endo-82-1-109. [DOI] [PubMed] [Google Scholar]
  24. Reichert L. E., Jr, Rasco M. A., Ward D. N., Niswender G. D., Midgley A. R., Jr Isolation and properties of subunits of bovine pituitary luteinizing hormone. J Biol Chem. 1969 Oct 10;244(19):5110–5117. [PubMed] [Google Scholar]
  25. Ryan R. J., Jiang N. S., Hanlon S. Physical properties of human follicle-stimulating hormone. Biochemistry. 1971 Apr 13;10(8):1321–1330. doi: 10.1021/bi00784a008. [DOI] [PubMed] [Google Scholar]
  26. STEELMAN S. L., POHLEY F. M. Assay of the follicle stimulating hormone based on the augmentation with human chorionic gonadotropin. Endocrinology. 1953 Dec;53(6):604–616. doi: 10.1210/endo-53-6-604. [DOI] [PubMed] [Google Scholar]
  27. Saxena B. B., Rathnam P. Purification of follicle-stimulating hormone from human pituitary glands. J Biol Chem. 1967 Sep 10;242(17):3769–3775. [PubMed] [Google Scholar]
  28. Sherwood O. D., Grimek H. J., McShan W. H. Purification and properties of follicle-stimulating hormone from sheep pituitary glands. J Biol Chem. 1970 May 10;245(9):2328–2336. [PubMed] [Google Scholar]
  29. Shiu R. P., Kelly P. A., Friesen H. G. Radioreceptor assay for prolactin and other lactogenic hormones. Science. 1973 Jun 1;180(4089):968–971. doi: 10.1126/science.180.4089.968. [DOI] [PubMed] [Google Scholar]
  30. Shome B., Parlow A. F. Human follicle stimulating hormone (hFSH): first proposal for the amino acid sequence of the alpha-subunit (hFSHa) and first demonstration of its identity with the alpha-subunit of human luteinizing hormone (hLHa). J Clin Endocrinol Metab. 1974 Jul;39(1):199–202. doi: 10.1210/jcem-39-1-199. [DOI] [PubMed] [Google Scholar]
  31. Shome B., Parlow A. F. Human follicle stimulating hormone: first proposal for the amino acid sequence of the hormone-specific, beta subunit (hFSHb). J Clin Endocrinol Metab. 1974 Jul;39(1):203–205. doi: 10.1210/jcem-39-1-203. [DOI] [PubMed] [Google Scholar]
  32. Tsushima T., Friesen H. G. Radioreceptor assay for growth hormone. J Clin Endocrinol Metab. 1973 Aug;37(2):334–337. doi: 10.1210/jcem-37-2-334. [DOI] [PubMed] [Google Scholar]
  33. Verrier B., Fayet G., Lissitzky S. Thyrotropin-binding properties of isolated thyroid cells and their purified plasma membranes. Relation of thyrotropin-specific binding to adenylate-cyclase activation. Eur J Biochem. 1974 Mar 1;42(2):355–365. doi: 10.1111/j.1432-1033.1974.tb03347.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES