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. 1982 Aug 1;205(2):295–302. doi: 10.1042/bj2050295

Cathepsin B from human renal cortex.

A D Gounaris, E E Slater
PMCID: PMC1158481  PMID: 7138503

Abstract

Cysteine-proteinase activity was observed in homogenates of human-cadaver renal cortex. This activity co-purified with renin enzymic activity until separation by aminohexyl-Sepharose--pepstatin affinity chromatography. The cysteine proteinase was purified 1780-fold after the following successive chromatographic procedures: Sephadex G-75, DEAE-cellulose DE-52, and an organomercurial affinity resin. The proteinase activity was dependent upon activation by thiol-containing compounds such as dithiothreitol, as well as by EDTA, and was inhibited by the thiol-group-specific alkylating reagents iodoacetic acid and N-ethylmaleimide. DE-52 cellulose chromatography resolved the cysteine proteinase into two components. On the basis of molecular size (26 000 daltons), activity as a function of pH, stability as a function of pH, substrate specificity and thermal lability, the major component (95%) has been identified as cathepsin B. The DE-52 cellulose elution pattern of the minor component (5%) is suggestive of cathepsin H [Schwartz & Barrett (1980) Biochem. J. 191, 487-497] Enzymic activity was determined with synthetic substrates, in particular alpha-N-benzoyl-DL-arginine 2-naphthylamide (Bz-Arg-NNap), thus precluding the detection of cathepsin L [Kirschke, Langner, Wiederanders, Ansorge, Bohley & Broghammer (1976) Acta Biol. Med. Germ. 35, 285-299]. Inhibition by dimethyl sulphoxide was observed in the determination of Km = 7.0 +/- 0.4 mM for the substrate Bz-Arg-NNap, and care must therefore be taken in the preparation of substrate solutions.

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Selected References

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  1. BLOSTEIN R., RUTTER W. J. COMPARATIVE STUDIES OF LIVER AND MUSCLE ALDOLASE. II. IMMUNOCHEMICAL AND CHROMATOGRAPHIC DIFFERENTIATION. J Biol Chem. 1963 Oct;238:3280–3285. [PubMed] [Google Scholar]
  2. Barrett A. J. A new assay for cathepsin B1 and other thiol proteinases. Anal Biochem. 1972 May;47(1):280–293. doi: 10.1016/0003-2697(72)90302-8. [DOI] [PubMed] [Google Scholar]
  3. Barrett A. J. An improved color reagent for use in Barrett's assay of Cathepsin B. Anal Biochem. 1976 Nov;76(50):374–376. doi: 10.1016/0003-2697(76)90298-0. [DOI] [PubMed] [Google Scholar]
  4. Barrett A. J. Human cathepsin B1. Purification and some properties of the enzyme. Biochem J. 1973 Apr;131(4):809–822. doi: 10.1042/bj1310809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bond J. S., Barrett A. J. Degradation of fructose-1,6-bisphosphate aldolase by cathepsin B. Biochem J. 1980 Jul 1;189(1):17–25. doi: 10.1042/bj1890017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Campbell P. N., Blobel G. The role of organelles in the chemical modification of the primary translation products of secretory proteins. FEBS Lett. 1976 Dec 31;72(2):215–226. doi: 10.1016/0014-5793(76)80973-8. [DOI] [PubMed] [Google Scholar]
  7. Distelmaier P., Hübner H., Otto K. Cathepsins B1 and B2 in various organs of the rat. Enzymologia. 1972 May 31;42(5):363–375. [PubMed] [Google Scholar]
  8. Evans P., Etherington D. J. Characterisation of cathepsin B and collagenolytic cathepsin from human placenta. Eur J Biochem. 1978 Feb 1;83(1):87–97. doi: 10.1111/j.1432-1033.1978.tb12071.x. [DOI] [PubMed] [Google Scholar]
  9. Gráf L., Kenessey A. Specific cleavage of a single peptide bond (residues 77-78) in beta-lipotropin by a pituitary endopeptidase. FEBS Lett. 1976 Oct 15;69(1):255–260. doi: 10.1016/0014-5793(76)80699-0. [DOI] [PubMed] [Google Scholar]
  10. Hardy M. F., Pennington R. J. Separation of cathepsin B1 and related enzymes from rat skeletal muscle. Biochim Biophys Acta. 1979 Apr 25;577(2):253–266. doi: 10.1016/0005-2795(79)90029-1. [DOI] [PubMed] [Google Scholar]
  11. Inagami T., Hirose S., Murakami K., Matoba T. Native form of renin in the kidney. J Biol Chem. 1977 Nov 10;252(21):7733–7737. [PubMed] [Google Scholar]
  12. Kirschke H., Langner J., Wiederanders B., Ansorge S., Bohley P., Broghammer U. Intrazellulärer Proteinabbau. VII. Kathepsin L und H: Zwei neue Proteinasen aus Rattenleberlysosomen. Acta Biol Med Ger. 1976;35(3-4):285–299. [PubMed] [Google Scholar]
  13. Knight C. G. Human cathepsin B. Application of the substrate N-benzyloxycarbonyl-L-arginyl-L-arginine 2-naphthylamide to a study of the inhibition by leupeptin. Biochem J. 1980 Sep 1;189(3):447–453. doi: 10.1042/bj1890447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. MacGregor R. R., Hamilton J. W., Shofstall R. E., Cohn D. V. Isolation and characterization of porcine parathyroid cathepsin B. J Biol Chem. 1979 Jun 10;254(11):4423–4427. [PubMed] [Google Scholar]
  16. Quinn P. S., Judah J. D. Calcium-dependent Golgi-vesicle fusion and cathepsin B in the conversion of proalbumin into albumin in rat liver. Biochem J. 1978 May 15;172(2):301–309. doi: 10.1042/bj1720301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schwartz W. N., Barrett A. J. Human cathepsin H. Biochem J. 1980 Nov 1;191(2):487–497. doi: 10.1042/bj1910487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Segal H. L. Mechanism and regulation of protein turnover in animal cells. Curr Top Cell Regul. 1976;11:183–201. doi: 10.1016/b978-0-12-152811-9.50012-2. [DOI] [PubMed] [Google Scholar]
  19. Singh H., Kalnitsky G. Separation of a new alpha-N-benzoylarginine-beta-naphthylamide hydrolase from cathepsin B1. Purification, characterization, and properties of both enzymes from rabbit lung. J Biol Chem. 1978 Jun 25;253(12):4319–4326. [PubMed] [Google Scholar]
  20. Slater E. E., Haber E. Inactive renin--"through a glass darkly". N Engl J Med. 1979 Aug 23;301(8):429–430. doi: 10.1056/NEJM197908233010810. [DOI] [PubMed] [Google Scholar]
  21. Slater E. E., Strout H. V., Jr Pure human renin. Identification and characterization and of two major molecular weight forms. J Biol Chem. 1981 Aug 10;256(15):8164–8171. [PubMed] [Google Scholar]
  22. TALLAN H. H., JONES M. E., FRUTON J. S. On the proteolytic enzymes of animal tissues. X. Beef spleen cathepsin C. J Biol Chem. 1952 Feb;194(2):793–805. [PubMed] [Google Scholar]
  23. Takahashi K., Isemura M., Ikenaka T. Isolation and characterization of three forms of cathepsin B from porcine liver. J Biochem. 1979 Apr;85(4):1053–1060. doi: 10.1093/oxfordjournals.jbchem.a132412. [DOI] [PubMed] [Google Scholar]
  24. Takahashi S., Murakami K., Miyake Y. Activation of kidney prorenin by kidney cathepsin B isozymes. J Biochem. 1982 Jan;91(1):419–422. doi: 10.1093/oxfordjournals.jbchem.a133705. [DOI] [PubMed] [Google Scholar]
  25. Towatari T., Katunuma N. Crystallization and amino acid composition of cathepsin B from rat liver lysosomes. Biochem Biophys Res Commun. 1978 Jul 28;83(2):513–520. doi: 10.1016/0006-291x(78)91020-3. [DOI] [PubMed] [Google Scholar]
  26. Towatari T., Kawabata Y., Katunuma N. Crystallization and properties of cathepsin B from rat liver. Eur J Biochem. 1979 Dec;102(1):279–289. doi: 10.1111/j.1432-1033.1979.tb06290.x. [DOI] [PubMed] [Google Scholar]
  27. Towatari T., Tanaka K., Yoshikawa D., Katunuma N. Purification and properties of a new cathepsin from rat liver. J Biochem. 1978 Sep;84(3):659–672. doi: 10.1093/oxfordjournals.jbchem.a132171. [DOI] [PubMed] [Google Scholar]
  28. Warwas M., Dobryszycka W. Cathepsins B1 from human fetal membranes. Biochim Biophys Acta. 1976 Apr 8;429(2):573–580. doi: 10.1016/0005-2744(76)90305-3. [DOI] [PubMed] [Google Scholar]
  29. Wootton R., Brereton P. J., Clark M. B., Hesp R., Hodkinson H. M., Klenerman L., Reeve J., Slavin G., Tellez-Yudilevich M. Fractured neck of femur in the elderly: an attempt to identify patients at risk. Clin Sci (Lond) 1979 Jul;57(1):93–101. doi: 10.1042/cs0570093. [DOI] [PubMed] [Google Scholar]

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