title: Affinity purification of the novel cysteine proteinase papaya proteinase IV, and papain from papaya latex.
abstract: A procedure is described for the purification of a previously undetected cysteine proteinase, which we have called papaya proteinase IV, from spray-dried latex of the papaya (Carica papaya) plant. The purification involves affinity chromatography on Gly-Phe-aminoacetonitrile linked to CH-Sepharose 4B, with elution by 2-hydroxyethyl disulphide at pH 4.5. The product thus obtained is a mixture of almost fully active papain and papay proteinase IV, which are then separated by cation-exchange chromatography. A preliminary characterization of papaya proteinase IV showed it to be very similar to chymopapain in both molecular size and charge. However, the new enzyme is immunologically distinct from the previously characterized cysteine proteinases of papaya latex. It also differs in its lack of activity against the synthetic substrates of the other papaya proteinases, in its narrow specificity against protein substrates and its lack of inhibition by chicken cystatin. Papaya proteinase IV is abundant, contributing almost 30% of the protein in spray-dried papaya latex, and contamination of chymopapain preparations with this enzyme may account for some of the previously reported heterogeneity of chymopapain.
authors.affiliation: Department of Biochemistry, Strangeways Research Laboratory, Cambridge, U.K.
authors.forename: D J
authors.initials: DJ
authors.lastname: Buttle
authors.forename: A A
authors.initials: AA
authors.lastname: Kembhavi
authors.forename: S L
authors.initials: SL
authors.lastname: Sharp
authors.forename: R E
authors.initials: RE
authors.lastname: Shute
authors.forename: D H
authors.initials: DH
authors.lastname: Rich
authors.forename: A J
authors.initials: AJ
authors.lastname: Barrett
mesh_terms: D002384:Catalysis; D002846:Chromatography, Affinity; D003546:Cysteine Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005779:Immunodiffusion; D007840:Latex / Q000302:isolation & purification*; D010206:Papain / Q000302:isolation & purification*; D010944:Plants / Q000201:enzymology
chemical_list: D007840:Latex; D003546:Cysteine Endopeptidases; D010206:Papain; C050874:glycyl endopeptidase