L-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) and Its Analogues As Inhibitors of Cysteine Proteinases Including Cathepsins B, H and L

Overview

Journal Biochem J

Specialty Biochemistry

Date 1982 Jan 1

PMID 7044372

Citations 249

Authors

A J Barrett

A A Kembhavi

M A Brown

H KIRSCHKE

C G Knight

M Tamai

K Hanada

Affiliations

Soon will be listed here.

Abstract

1. L-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) at a concentration of 0.5 mM had no effect on the serine proteinases plasma kallikrein and leucocyte elastase or the metalloproteinases thermolysin and clostridial collagenase. In contrast, 10 muM-E-64 rapidly inactivated the cysteine proteinases cathepsins B, H and L and papain (t0.5 = 0.1-17.3s). The streptococcal cysteine proteinase reacted much more slowly, and there was no irreversible inactivation of clostripain. The cysteine-dependent exopeptidase dipeptidyl peptidase I was very slowly inactivated by E-64. 2. the active-site-directed nature of the interaction of cathepsin B and papain with E-64 was established by protection of the enzyme in the presence of the reversible competitive inhibitor leupeptin and by the stereospecificity for inhibition by the L as opposed to the D compound. 3. It was shown that the rapid stoichiometric reaction of the cysteine proteinases related to papain can be used to determine the operational molarity of solutions of the enzymes and thus to calibrate rate assays. 4. The apparent second-order rate constants for the inactivation of human cathepsins B and H and rat cathepsin L by a series of structural analogues of E-64 are reported, and compared with those for some other active-site-directed inhibitors of cysteine proteinases. 5. L-trans-Epoxysuccinyl-leucylamido(3-methyl)butane (Ep-475) was found to inhibit cathepsins B and L more rapidly than E-64. 6. Fumaryl-leucylamido(3-methyl)butane (Dc-11) was 100-fold less reactive than the corresponding epoxide, but was nevertheless about as effective as iodoacetate.

Citing Articles

Enteropathogenic E. coli effector Map interacts with Rab13 and regulates the depletion of the tight junction proteins occludin and claudins via cathepsin B-mediated mechanisms.

Mandal A, Walling P, Qureshi S, Kansal K, Aijaz S Biol Open. 2025; 14(2).

PMID: 39912222 PMC: 11892358. DOI: 10.1242/bio.061794.

Activation of VGLL4 Suppresses Cardiomyocyte Maturational Hypertrophic Growth.

Farley A, Gao Y, Sun Y, Zohrabian S, Pu W, Lin Z Cells. 2024; 13(16.

PMID: 39195232 PMC: 11352427. DOI: 10.3390/cells13161342.

Cathepsins L and B target HIF1α for oxygen-independent proteolytic cleavage.

Stuart S, Tarade D, Ohh M Sci Rep. 2024; 14(1):14799.

PMID: 38926538 PMC: 11208597. DOI: 10.1038/s41598-024-65537-9.

Nature-Inspired Gallinamides Are Potent Antischistosomal Agents: Inhibition of the Cathepsin B1 Protease Target and Binding Mode Analysis.

Spiwokova P, Horn M, Fanfrlik J, Jilkova A, Fajtova P, Leontovyc A ACS Infect Dis. 2024; 10(6):1935-1948.

PMID: 38757505 PMC: 11184554. DOI: 10.1021/acsinfecdis.3c00589.

Structural Elucidation and Antiviral Activity of Covalent Cathepsin L Inhibitors.

Falke S, Lieske J, Herrmann A, Loboda J, Karnicar K, Gunther S J Med Chem. 2024; 67(9):7048-7067.

PMID: 38630165 PMC: 11089505. DOI: 10.1021/acs.jmedchem.3c02351.

References

Saklatvala J, Barrett A . Identification of proteinases in rheumatoid synovium. Detection of leukocyte elastase cathepsin G and another serine proteinase. Biochim Biophys Acta. 1980; 615(1):167-77. DOI: 10.1016/0005-2744(80)90020-0. View

Takio K, Towatari T, Katunuma N, Titani K . Primary structure study of rat liver cathepsin B -- a striking resemblance to papain. Biochem Biophys Res Commun. 1980; 97(1):340-6. DOI: 10.1016/s0006-291x(80)80173-2. View

Hashida S, Towatari T, Kominami E, Katunuma N . Inhibitions by E-64 derivatives of rat liver cathepsin B and cathepsin L in vitro and in vivo. J Biochem. 1980; 88(6):1805-11. DOI: 10.1093/oxfordjournals.jbchem.a133155. View

Green G, Shaw E . Peptidyl diazomethyl ketones are specific inactivators of thiol proteinases. J Biol Chem. 1981; 256(4):1923-8. View

Knight C . 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; 189(3):447-53. PMC: 1162023. DOI: 10.1042/bj1890447. View

Schwartz W, Barrett A . Human cathepsin H. Biochem J. 1980; 191(2):487-97. PMC: 1162239. DOI: 10.1042/bj1910487. View

Tamai M, Hanada K, Adachi T, Oguma K, Kashiwagi K, Omura S . Papain inhibitions by optically active E-64 analogs. J Biochem. 1981; 90(1):255-7. DOI: 10.1093/oxfordjournals.jbchem.a133458. View

Nagase H, Barrett A . Human plasma kallikrein. A rapid purification method with high yield. Biochem J. 1981; 193(1):187-92. PMC: 1162589. DOI: 10.1042/bj1930187. View

KIRSCHKE H, Shaw E . Rapid interaction of cathepsin L by Z-Phe-PheCHN12 and Z-Phe-AlaCHN2. Biochem Biophys Res Commun. 1981; 101(2):454-8. DOI: 10.1016/0006-291x(81)91281-x. View

10.

Barrett A . Human cathepsin B1. Purification and some properties of the enzyme. Biochem J. 1973; 131(4):809-22. PMC: 1177541. DOI: 10.1042/bj1310809. View

11.

Brocklehurst K, Little G . Reactions of papain and of low-molecular-weight thiols with some aromatic disulphides. 2,2'-Dipyridyl disulphide as a convenient active-site titrant for papain even in the presence of other thiols. Biochem J. 1973; 133(1):67-80. PMC: 1177671. DOI: 10.1042/bj1330067. View

12.

Thompson R, BLOUT E . Peptide chloromethyl ketones as irreversible inhibitors of elastase. Biochemistry. 1973; 12(1):44-7. DOI: 10.1021/bi00725a008. View

13.

Kortt A, Liu T . On the mechanism of action of streptococcal proteinase. II. Comparison of the kinetics of proteinase- and papain-catalyzed hydrolysis of N-acylamino acid esters. Biochemistry. 1973; 12(2):328-37. DOI: 10.1021/bi00726a024. View

14.

Berger A, Schechter I . Mapping the active site of papain with the aid of peptide substrates and inhibitors. Philos Trans R Soc Lond B Biol Sci. 1970; 257(813):249-64. DOI: 10.1098/rstb.1970.0024. View

15.

Bender M, BEGUE-CANTON M, Blakeley R, Brubacher L, Feder J, GUNTER C . The determination of the concentration of hydrolytic enzyme solutions: alpha-chymotrypsin, trypsin, papain, elastase, subtilisin, and acetylcholinesterase. J Am Chem Soc. 1966; 88(24):5890-913. DOI: 10.1021/ja00976a034. View

16.

Gerwin B, STEIN W, Moore S . On the specificity of streptococcal proteinase. J Biol Chem. 1966; 241(14):3331-9. View

17.

Mort J, Recklies A, Poole A . Characterization of a thiol proteinase secreted by malignant human breast tumours. Biochim Biophys Acta. 1980; 614(1):134-43. DOI: 10.1016/0005-2744(80)90174-6. View

18.

Sugita H, Ishiura S, Suzuki K, Imahori K . Inhibition of epoxide derivatives on chicken calcium-activated neutral protease (CANP) in vitro and in vivo. J Biochem. 1980; 87(1):339-41. DOI: 10.1093/oxfordjournals.jbchem.a132742. View

19.

Towatari T, Tanaka K, Yoshikawa D, Katunuma N . Purification and properties of a new cathepsin from rat liver. J Biochem. 1978; 84(3):659-72. DOI: 10.1093/oxfordjournals.jbchem.a132171. View

20.

Emod I, Keil B . Five Sepharose-bound ligands for the chromatographic purification of Clostridium collagenase and clostripain. FEBS Lett. 1977; 77(1):51-6. DOI: 10.1016/0014-5793(77)80191-9. View