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Current Drug Discovery Technologies

Editor-in-Chief

ISSN (Print): 1570-1638
ISSN (Online): 1875-6220

The Characterisation of the Collagenolytic Activity of Cardosin A Demonstrates its Potential Application for Extracellular Matrix Degradative Processes

Author(s): Ana S. Duarte, Anabela O. Pereira, Antonio M. S. Cabrita, Arthur J. G. Moir, Euclides M. V. Pires and Marlene M. T. Barros

Volume 2, Issue 1, 2005

Page: [37 - 44] Pages: 8

DOI: 10.2174/1570163053175448

Price: $65

Abstract

Type I collagen is the major fibrous protein of mammals being needed to strengthen and organise the extracellular matrix (ECM). Connective tissue components are modulated by matrix metalloproteinases, which are critical for disintegration and remodelling of ECM under physiological and pathological conditions. Cardosin A is an abundant aspartic proteinase (AP) from Cynara cardunculus L. that has been shown to be able to hydrolyse fibrillar collagen within the α-chains. The aim of this work is the characterisation of collagen degradation by cardosin A, since in the native state fibrillar collagen is resistant to most proteolytic enzymes. The pattern of type I collagen hydrolysis by cardosin A is defined and maintained for at least 24 hours of digestion, suggesting that cardosin A can hydrolyse collagen at a small number of specific peptide bonds. N-terminal sequencing of hydrolysis products identified one cleavage site as being Phe464-Gln465 in the a2 chain of collagen I. Two peptides were synthesised correspondent to collagen I specific sequences, in order to produce two polyclonal antibodies, that allowed the identification of three collagen fragments following cardosin A cleavage. Defining the mechanism of collagen cleavage by collagenases and other enzymes, like cardosin A, is important for the comprehension of physiological and pathological processes affecting the ECM. To our knowledge, this is the first study of in vitro collagenolytic activity of a plant AP. Therefore, in view of the cardosin A restricted specificity towards collagen this enzyme may be proposed for an eventual medical or technical procedures assisting ECM remodelling.

Keywords: type I collagen, cardosin a, aspartic proteinase, fibrosis, adhesion


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