Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1983 May 1;211(2):519–522. doi: 10.1042/bj2110519

Purification of human renin by affinity chromatography using a new peptide inhibitor of renin, H.77 (D-His-Pro-Phe-His-LeuR-Leu-Val-Tyr).

G D McIntyre, B Leckie, A Hallett, M Szelke
PMCID: PMC1154389  PMID: 6347189

Abstract

A new affinity column for renin was prepared by coupling the isosteric peptide inhibitor of renin, H.77 (D-His-Pro-Phe-His-LeuR-Leu-Val-Tyr, where R is a reduced isosteric bond, -CH2-NH-), to activated 6-aminohexanoic acid-Sepharose 4B. Chromatography of a crude extract of human kidney cortex on this material resulted in a 5500-fold purification of renin in 76% yield. The purified enzyme (specific activity 871 units/mg) was free of non-specific acid-proteinase activity and was stable at pH 6.8 and -20 degrees C over a period of several weeks.

Full text

PDF
519

Images in this article

521Fig. 2.
on p.521

Selected References

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

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Böhlen P., Stein S., Dairman W., Udenfriend S. Fluorometric assay of proteins in the nanogram range. Arch Biochem Biophys. 1973 Mar;155(1):213–220. doi: 10.1016/s0003-9861(73)80023-2. [DOI] [PubMed] [Google Scholar]
  3. Dubin A., Suen E. T., Delaney R., Chiu A., Johnson B. C. Regulation of vitamin K-dependent carboxylation. J Biol Chem. 1980 Jan 25;255(2):349–352. [PubMed] [Google Scholar]
  4. Galen F. X., Devaux C., Guyenne T., Menard J., Corvol P. Multiple forms of human renin. Purification and characterization. J Biol Chem. 1979 Jun 10;254(11):4848–4855. [PubMed] [Google Scholar]
  5. HAAS E., LAMFROM H., GOLDBLATT H. A simple method for the extraction and partial purification of renin. Arch Biochem Biophys. 1954 Feb;48(2):256–260. doi: 10.1016/0003-9861(54)90339-2. [DOI] [PubMed] [Google Scholar]
  6. Hille M. B., Barrett A. J., Dingle J. T., Fell H. B. Microassay for cathepsin D shows an unexpected effedt of cycloheximide on limb-bone rudiments in organ culture. Exp Cell Res. 1970 Aug;61(2):470–472. doi: 10.1016/0014-4827(70)90476-3. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Macart M., Gerbaut L. An improvement of the Coomassie Blue dye binding method allowing an equal sensitivity to various proteins: application to cerebrospinal fluid. Clin Chim Acta. 1982 Jun 16;122(1):93–101. doi: 10.1016/0009-8981(82)90100-0. [DOI] [PubMed] [Google Scholar]
  9. Millar J. A., Leckie B. J., Morton J. J., Jordan J., Tree M. A microassay for active and total renin concentration in human plasma based on antibody trapping. Clin Chim Acta. 1980 Feb 14;101(1):5–15. doi: 10.1016/0009-8981(80)90050-9. [DOI] [PubMed] [Google Scholar]
  10. Peart W. S., Lloyd A. M., Thatcher G. N., Lever A. F., Payne N., Stone N. Purification of pig renin. Biochem J. 1966 Jun;99(3):708–716. doi: 10.1042/bj0990708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Righetti P. G., Caravaggio T. Isoelectric points and molecular weights of proteins. J Chromatogr. 1976 Apr 21;127(11):1–28. doi: 10.1016/s0021-9673(00)98537-6. [DOI] [PubMed] [Google Scholar]
  12. Slater E. E. Renin. Methods Enzymol. 1981;80(Pt 100):427–442. doi: 10.1016/s0076-6879(81)80037-7. [DOI] [PubMed] [Google Scholar]
  13. Szelke M., Leckie B. J., Tree M., Brown A., Grant J., Hallett A., Hughes M., Jones D. M., Lever A. F. H-77: a potent new renin inhibitor. In vitro and in vivo studies. Hypertension. 1982 May-Jun;4(3 Pt 2):59–69. [PubMed] [Google Scholar]
  14. Szelke M., Leckie B., Hallett A., Jones D. M., Sueiras J., Atrash B., Lever A. F. Potent new inhibitors of human renin. Nature. 1982 Oct 7;299(5883):555–557. doi: 10.1038/299555a0. [DOI] [PubMed] [Google Scholar]
  15. Wray W., Boulikas T., Wray V. P., Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. doi: 10.1016/0003-2697(81)90179-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES