Chymopapain. Chromatographic Purification and Immunological Characterization

Overview

Journal Biochem J

Specialty Biochemistry

Date 1984 Oct 1

PMID 6437389

Citations 15

Authors

D J Buttle

A J Barrett

Affiliations

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Abstract

Chymopapain (EC 3.4.22.6) was purified from commercially available spray-dried latex of papaya (Carica papaya) fruit by (NH4)2SO4 fractionation and fast protein chromatography on the Mono S cation-exchange column. Multiple forms of chymopapain separated chromatographically were shown to be immunologically identical. A major form was isolated and found to be homogeneous by several criteria, and fully active, and its N-terminal amino acid was identified as tyrosine. Latex from fresh unripe papaya fruit contained predominantly one form of chymopapain, and it is concluded that chymopapain is a single enzyme distinct from the other cysteine proteinases of C. papaya latex.

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References

Kunimitsu D, Yasunobu K . Chymopapain. IV. The chromatographic fractionation of partially purified chymopapain and the characterization of crystalline chymopapain B. Biochim Biophys Acta. 1967; 139(2):405-17. DOI: 10.1016/0005-2744(67)90044-7. View

Schack P . Fractionation of proteolytic enzymes of dried papaya latex. Isolation and preliminary characterization of a new proteolytic enzyme. C R Trav Lab Carlsberg. 1967; 36(4):67-83. View

LYNN K . An isolation of chymopapain. J Chromatogr. 1973; 84(2):423-5. DOI: 10.1016/s0021-9673(01)91729-7. View

Clagett J, Tokuda S, ENGELHARD W . Chymopapain C, an immunosuppressive protease. I. Partial purification and characterization. Proc Soc Exp Biol Med. 1974; 145(4):1250-7. DOI: 10.3181/00379727-145-37991. View

Barrett A . Chicken alpha2-proteinase inhibitor: a serum protein homologous with ovoinhibitor of egg white. Biochim Biophys Acta. 1974; 371(1):52-62. DOI: 10.1016/0005-2795(74)90154-8. View

Robinson G . Isolation and characterization of papaya peptidase A from commercial chymopapain. Biochemistry. 1975; 14(16):3695-700. DOI: 10.1021/bi00687a028. View

Drenth J, Kalk K, Swen H . Binding of chloromethyl ketone substrate analogues to crystalline papain. Biochemistry. 1976; 15(17):3731-8. DOI: 10.1021/bi00662a014. View

Baines B, 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; 177(2):541-8. PMC: 1186404. DOI: 10.1042/bj1770541. View

LYNN K . A purification and some properties of two proteases from papaya latex. Biochim Biophys Acta. 1979; 569(2):193-201. DOI: 10.1016/0005-2744(79)90054-8. View

10.

Brocklehurst K, Baines B, Mushiri M . Evidence that the active centre of chymopapain A is different from the active centres of some other cysteine proteinases and that the Brønsted coefficient (beta nuc.) for the reactions of thiolate anions with 2,2'-dipyridyl disulphide may be.... Biochem J. 1980; 189(1):189-29. PMC: 1161932. DOI: 10.1042/bj1890189. View

11.

Polgar L . Isolation of highly active papaya peptidases A and B from commercial chymopapain. Biochim Biophys Acta. 1981; 658(2):262-9. DOI: 10.1016/0005-2744(81)90296-5. View

12.

Barrett A, KIRSCHKE H . Cathepsin B, Cathepsin H, and cathepsin L. Methods Enzymol. 1981; 80 Pt C:535-61. DOI: 10.1016/s0076-6879(81)80043-2. View

13.

Barrett A, Kembhavi A, Brown M, KIRSCHKE H, Knight C, Tamai M . 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; 201(1):189-98. PMC: 1163625. DOI: 10.1042/bj2010189. View

14.

JAVID M, Nordby E, Ford L, Hejna W, Whisler W, Burton C . Safety and efficacy of chymopapain (Chymodiactin) in herniated nucleus pulposus with sciatica. Results of a randomized, double-blind study. JAMA. 1983; 249(18):2489-94. View

15.

Brocklehurst K, Salih E . A re-evaluation of the nomenclature of the cysteine proteinases of Carica papaya and a rational basis for their identification. Biochem J. 1983; 213(2):559-60. PMC: 1152164. DOI: 10.1042/bj2130559. View

16.

Ebata M, Yasunobu K . Chymopapain. I. Isolation, crystallization, and preliminary characterization. J Biol Chem. 1962; 237:1086-94. View