Two new cysteine endopeptidases obtained from the latex of Araujia hortorum fruits

Two new endopeptidases were purified to homogeneity from the latex of Araujia hortorum fruits by a simple purification procedure involving ultracentrifugation and ion exchange chromatography. Molecular weights of araujiain h II and araujiain h III were 23,718 and 23546 (mass spectrometry), respectiv...

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Bibliographic Details
Authors: Obregón, Walter David, Arribére, María Cecilia, Morcelle del Valle, Susana Raquel, Liggieri, Constanza Silvina, Caffini, Néstor Oscar, Priolo de Lufrano, Nora Silvia
Format: article
Status:Published version
Publication Date:2001
Country:Argentina
Institution:Universidad Nacional de La Plata
Repository:SEDICI (UNLP)
Language:English
OAI Identifier:oai:sedici.unlp.edu.ar:10915/143564
Online Access:http://sedici.unlp.edu.ar/handle/10915/143564
Access Level:Open access
Keyword:Química
Araujiain
Cysteine endopeptidases
Latex
Milkweed family
Protein purification
Description
Summary:Two new endopeptidases were purified to homogeneity from the latex of Araujia hortorum fruits by a simple purification procedure involving ultracentrifugation and ion exchange chromatography. Molecular weights of araujiain h II and araujiain h III were 23,718 and 23546 (mass spectrometry), respectively. The isoelectric point of araujiain h II was 8.9, whereas araujiain h III had a pI higher than 9.3. Maximum proteolytic activity on caseine was reached at pH 8.0-9.0 for both endopeptidases, which were irreversibly inhibited by iodoacetate and E-64, suggesting they belong to the cysteine protease family. Esterolytic activity was determined on N-alpha-CBZ-amino acid-p-nitrophenyl esters, and the highest kcat/Km values for the both enzymes were obtained with the glutamine derivative. The N-terminal sequences of araujiain h II and araujiain h III showed a high degree of homology with other plant cysteine endopeptidases.