Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1984 Mar 15;218(3):939–946. doi: 10.1042/bj2180939

Cystatin-like cysteine proteinase inhibitors from human liver.

G D Green, A A Kembhavi, M E Davies, A J Barrett
PMCID: PMC1153426  PMID: 6426465

Abstract

Cysteine proteinase inhibitor (CPI) forms from human liver were purified from the tissue homogenate by alkaline denaturation of cysteine proteinases with which they are complexed, acetone fractionation, affinity chromatography on S-carboxymethyl-papain-Sepharose and chromatofocusing. The multiple forms of CPI were shown immunologically to be forms of two proteins, referred to as CPI-A (comprising the forms of relatively acidic pI) and CPI-B (comprising the more basic forms). CPI-A and CPI-B are similar in their Mr of about 12400, considerable stability to pH2, pH11 and 80 degrees C, and tight-binding inhibition of papain, several related cysteine proteinases and dipeptidyl peptidase I. Ki values were determined for papain, human cathepsins B, H and L, and dipeptidyl peptidase I. The affinity of CPI-A for cathepsin B was about 10-fold greater than that of CPI-B, whereas CBI-B showed about 100-fold stronger inhibition of dipeptidyl peptidase I. For all the cysteine proteinases the liver inhibitors were somewhat less tight binding than cystatin. The resemblance of both CPI-A and CPI-B in several respects to egg-white cystatin is discussed. CPI-A seems to correspond to the epithelial inhibitor described previously, and CPI-B to the inhibitor from other cell types [Järvinen & Rinne (1982) Biochim. Biophys. Acta 708, 210-217].

Full text

PDF
939

Images in this article

943Fig. 2.
on p.943

Selected References

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

  1. Anastasi A., Brown M. A., Kembhavi A. A., Nicklin M. J., Sayers C. A., Sunter D. C., Barrett A. J. Cystatin, a protein inhibitor of cysteine proteinases. Improved purification from egg white, characterization, and detection in chicken serum. Biochem J. 1983 Apr 1;211(1):129–138. doi: 10.1042/bj2110129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baines B. S., Brocklehurst K. A necessary modification to the preparation of papain from any high-quality latex of Carica papaya and evidence for the structural integrity of the enzyme produced by traditional methods. Biochem J. 1979 Feb 1;177(2):541–548. doi: 10.1042/bj1770541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barrett A. J. Cathepsin D. Purification of isoenzymes from human and chicken liver. Biochem J. 1970 Apr;117(3):601–607. doi: 10.1042/bj1170601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barrett A. J. Chicken alpha2-proteinase inhibitor: a serum protein homologous with ovoinhibitor of egg white. Biochim Biophys Acta. 1974 Nov 5;371(1):52–62. doi: 10.1016/0005-2795(74)90154-8. [DOI] [PubMed] [Google Scholar]
  5. Barrett A. J., Kembhavi A. A., Brown M. A., Kirschke H., Knight C. G., Tamai M., Hanada K. L-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) and its analogues as inhibitors of cysteine proteinases including cathepsins B, H and L. Biochem J. 1982 Jan 1;201(1):189–198. doi: 10.1042/bj2010189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barrett A. J., Kirschke H. Cathepsin B, Cathepsin H, and cathepsin L. Methods Enzymol. 1981;80(Pt 100):535–561. doi: 10.1016/s0076-6879(81)80043-2. [DOI] [PubMed] [Google Scholar]
  7. Brzin J., Kopitar M., Locnikar P., Turk V. An endogenous inhibitor of cysteine and serine proteinases from spleen. FEBS Lett. 1982 Feb 22;138(2):193–197. doi: 10.1016/0014-5793(82)80439-0. [DOI] [PubMed] [Google Scholar]
  8. Bürk R. R., Eschenbruch M., Leuthard P., Steck G. Sensitive detection of proteins and peptides in polyacrylamide gels after formaldehyde fixation. Methods Enzymol. 1983;91:247–254. doi: 10.1016/s0076-6879(83)91021-2. [DOI] [PubMed] [Google Scholar]
  9. Coombs G. H. Proteinases of Leishmania mexicana and other flagellate protozoa. Parasitology. 1982 Feb;84(1):149–155. doi: 10.1017/s003118200005174x. [DOI] [PubMed] [Google Scholar]
  10. Dayhoff M. O., Barker W. C., Hunt L. T. Establishing homologies in protein sequences. Methods Enzymol. 1983;91:524–545. doi: 10.1016/s0076-6879(83)91049-2. [DOI] [PubMed] [Google Scholar]
  11. Dresden M. H., Payne D. C., Basch P. F. Acidic thiol proteinase activity of Schistosoma mansoni cultured in vitro. Mol Biochem Parasitol. 1982 Oct;6(4):203–208. doi: 10.1016/0166-6851(82)90054-8. [DOI] [PubMed] [Google Scholar]
  12. Gauthier F., Pagano M., Esnard F., Mouray H., Engler R. A heat stable low molecular weight inhibitor of lysosomal cysteine proteinases in human serum. Biochem Biophys Res Commun. 1983 Jan 27;110(2):449–455. doi: 10.1016/0006-291x(83)91170-1. [DOI] [PubMed] [Google Scholar]
  13. Henderson P. J. A linear equation that describes the steady-state kinetics of enzymes and subcellular particles interacting with tightly bound inhibitors. Biochem J. 1972 Apr;127(2):321–333. doi: 10.1042/bj1270321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hibino T., Fukuyama K., Epstein W. L. Chemical characterization, synthesis and distribution of proteinase inhibitor in newborn rat epidermis. Biochim Biophys Acta. 1980 Oct 1;632(2):214–226. doi: 10.1016/0304-4165(80)90079-3. [DOI] [PubMed] [Google Scholar]
  15. Hibino T., Fukuyama K., Epstein W. L. In vitro and in vivo inhibition of rat liver cathepsin L by epidermal proteinase inhibitor. Biochem Biophys Res Commun. 1980 Mar 28;93(2):440–447. doi: 10.1016/0006-291x(80)91097-9. [DOI] [PubMed] [Google Scholar]
  16. Hirado M., Iwata D., Niinobe M., Fujii S. Purification and properties of thiol protease inhibitor from rat liver cytosol. Biochim Biophys Acta. 1981 Jun 29;669(1):21–27. doi: 10.1016/0005-2795(81)90218-x. [DOI] [PubMed] [Google Scholar]
  17. Järvinen M. Purification and some characteristics of the human epidermal SH-protease inhibitor. J Invest Dermatol. 1978 Aug;71(2):114–118. doi: 10.1111/1523-1747.ep12546165. [DOI] [PubMed] [Google Scholar]
  18. Järvinen M., Rinne A. Human spleen cysteineproteinase inhibitor. Purification, fractionation into isoelectric variants and some properties of the variants. Biochim Biophys Acta. 1982 Nov 9;708(2):210–217. [PubMed] [Google Scholar]
  19. Järvinen M., Räsänen O., Rinne A. The low-molecular-weight SH-protease inhibitor in rat skin is epidermal. J Invest Dermatol. 1978 Aug;71(2):119–121. doi: 10.1111/1523-1747.ep12546254. [DOI] [PubMed] [Google Scholar]
  20. Kominami E., Wakamatsu N., Katunuma N. Endogenous thiol protease inhibitor from rat liver. Biochem Biophys Res Commun. 1981 Mar 31;99(2):568–575. doi: 10.1016/0006-291x(81)91783-6. [DOI] [PubMed] [Google Scholar]
  21. Kominami E., Wakamatsu N., Katunuma N. Purification and characterization of thiol proteinase inhibitor from rat liver. J Biol Chem. 1982 Dec 25;257(24):14648–14652. [PubMed] [Google Scholar]
  22. Kopitar M., Brzin J., Locnikar P., Turk V. Inhibitory mechanism of sericystatin, an intracellular proteinase inhibitor, reacting with cysteine proteinases. Hoppe Seylers Z Physiol Chem. 1981 Oct;362(10):1411–1414. [PubMed] [Google Scholar]
  23. Lenney J. F., Tolan J. R., Sugai W. J., Lee A. G. Thermostable endogenous inhibitors of cathepsins B and H. Eur J Biochem. 1979 Nov 1;101(1):153–161. doi: 10.1111/j.1432-1033.1979.tb04227.x. [DOI] [PubMed] [Google Scholar]
  24. Machleidt W., Borchart U., Fritz H., Brzin J., Ritonja A., Turk V. Protein inhibitors of cysteine proteinases. II. Primary structure of stefin, a cytosolic protein inhibitor of cysteine proteinases from human polymorphonuclear granulocytes. Hoppe Seylers Z Physiol Chem. 1983 Nov;364(11):1481–1486. doi: 10.1515/bchm2.1983.364.2.1481. [DOI] [PubMed] [Google Scholar]
  25. Rinne A., Järvinen M., Räsänen O., Dorn A. Nachweis eines epidermalen SH-Protease-Inhibitors in normalen Epithelien un einigen Neoplasmen beim Menschen--eine immunologische Studie. Acta Histochem Suppl. 1980;22:325–329. [PubMed] [Google Scholar]
  26. Robinson G. W. Isolation and characterization of papaya peptidase A from commercial chymopapain. Biochemistry. 1975 Aug 12;14(16):3695–3700. doi: 10.1021/bi00687a028. [DOI] [PubMed] [Google Scholar]
  27. Roughley P. J., Murphy G., Barrett A. J. Proteinase inhibitors of bovine nasal cartilage. Biochem J. 1978 Mar 1;169(3):721–724. doi: 10.1042/bj1690721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Saklatvala J., Barrett A. J. Identification of proteinases in rheumatoid synovium. Detection of leukocyte elastase cathepsin G and another serine proteinase. Biochim Biophys Acta. 1980 Sep 9;615(1):167–177. doi: 10.1016/0005-2744(80)90020-0. [DOI] [PubMed] [Google Scholar]
  29. Schwabe C., Anastasi A., Crow H., McDonald J. K., Barrett A. J. Cystatin. Amino acid sequence and possible secondary structure. Biochem J. 1984 Feb 1;217(3):813–817. doi: 10.1042/bj2170813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Takio K., Kominami E., Wakamatsu N., Katunuma N., Titani K. Amino acid sequence of rat liver thiol proteinase inhibitor. Biochem Biophys Res Commun. 1983 Sep 30;115(3):902–908. doi: 10.1016/s0006-291x(83)80020-5. [DOI] [PubMed] [Google Scholar]
  32. UDAKA K., HAYASHI H. FURTHER PURIFICATION OF A PROTEASE INHIBITOR FROM RABBIT SKIN WITH HEALING INFLAMMATION. Biochim Biophys Acta. 1965 Feb 15;97:251–261. doi: 10.1016/0304-4165(65)90089-9. [DOI] [PubMed] [Google Scholar]
  33. WILKINSON G. N. Statistical estimations in enzyme kinetics. Biochem J. 1961 Aug;80:324–332. doi: 10.1042/bj0800324. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES