Benzyloxycarbonylphenylalanylcitrulline p-nitroanilide as a substrate for papain and other plant cysteine proteinases.

Biochem J. 1984 April 1; 219(1): 325–328.

PMCID: PMC1153481

C J Gray, J Boukouvalas, R J Szawelski, and C W Wharton

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Abstract

After preliminary assays, with papain, bromelain and ficin, on a range of citrulline p-nitroanilides, values of Km and kcat. for the papain-catalysed hydrolysis of three derivatives, N alpha- benzyloxycarbonylcitrulline p-nitroanilide, benzyloxycarbonylphenylalanylcitrulline p-nitroanilide and benzyloxycarbonylglycylphenylalanylcitrulline p-nitroanilide, were obtained. It is concluded that benzyloxycarbonylphenylalanylcitrulline p-nitroanilide is a highly selective substrate for the sensitive detection and assay of the plant cysteine proteinases.

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Angelides, KJ; Fink, AL. Mechanism of Action of papain with a specific anilide substrate. Biochemistry. 1979 May 29;18(11):2355–2363. [PubMed]
Baines, BS; Brocklehurst, K. A spectrophotometric method for the detection of contaminant chymopapains in preparations of papain. Selective modification of one type of thiol group in the chymopapains by a two-protonic-state reagent. Biochem J. 1978 Jul 1;173(1):345–347. [PubMed]
Baines, BS; 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. [PubMed]
Brubacher, LJ; Bender, ML. The preparation and properties of trans-cinnamoyl-papain. J Am Chem Soc. 1966 Dec 20;88(24):5871–5880. [PubMed]
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 Feb 12;257(813):249–264. [PubMed]
Brubacher, LJ; Bender, ML. The reaction of trans-cinnamoyl-papain with a series of polyglycinamides of varying chain length. Biochem Biophys Res Commun. 1967 Apr 20;27(2):176–182. [PubMed]
Bruton, CJ; Hartley, BS. Chemical studies on methionyl-tRNA synthetase from Escherichia coli. J Mol Biol. 1970 Sep 14;52(2):165–178. [PubMed]
Leary, R; Larsen, D; Watanabe, H; Shaw, E. Diazomethyl ketone substrate derivatives as active-site-directed inhibitors of thiol proteases. Papain. Biochemistry. 1977 Dec 27;16(26):5857–5861. [PubMed]
Lowe, G; Yuthavong, Y. Kinetic specificity in papain-catalysed hydrolyses. Biochem J. 1971 Aug;124(1):107–115. [PubMed]
Mole, JE; Horton, HR. Kinetics of papain-catalyzed hydrolysis of -N-benzoyl-L-arginine-p-nitroanilide. Biochemistry. 1973 Feb 27;12(5):816–822. [PubMed]
Polgár, L. Deuterium isotope effects on papain acylation. Evidence for lack of general base catalysis and for enzyme--leaving-group interaction. Eur J Biochem. 1979 Aug 1;98(2):369–374. [PubMed]
Polgár, L; Halász, P. Evidence for multiple reactive forms of papain. Eur J Biochem. 1978 Aug 1;88(2):513–521. [PubMed]
Schechter, I; Berger, A. On the size of the active site in proteases. I. Papain. Biochem Biophys Res Commun. 1967 Apr 20;27(2):157–162. [PubMed]
Schechter, I; Berger, A. On the active site of proteases. 3. Mapping the active site of papain; specific peptide inhibitors of papain. Biochem Biophys Res Commun. 1968 Sep 6;32(5):898–902. [PubMed]
Stuchbury, T; Shipton, M; Norris, R; Malthouse, JP; Brocklehurst, K; Herbert, JA; Suschitzky, H. A reporter group delivery system with both absolute and selective specificity for thiol groups and an improved fluorescent probe containing the 7-nitrobenzo-2-oxa-1,3-diazole moiety. Biochem J. 1975 Nov;151(2):417–432. [PubMed]
Tokura, S; Nishi, N; Noguchi, J. A new substrate for papain, benzoyl-L-arginine-p-nitroanilide (L-BAPA). J Biochem. 1971 Mar;69(3):599–600. [PubMed]
Wharton, CW; Cornish-Bowden, A; Brocklehurst, K; Crook, EM. Kinetics of the hydrolysis of N-benzoyl-L-serine methyl ester catalysed by bromelain and by papain. Analysis of modifier mechanisms by lattice nomography, computational methods of parameter evaluation for substrate-activated catalyses and consequences of postulated non-productive binding in bromelain- and papain-catalysed hydrolyses. Biochem J. 1974 Aug;141(2):365–381. [PubMed]