IngentaConnect Degradation of extracellular matrix components by defined protein...

Abstract:

Summary Background

Larvae of the greenbottle fly Lucilia sericata are used routinely for the clinical treatment of difficult necrotic and infected wounds. Degradation by proteinases contained in larval excretory/secretory (ES) products is thought to contribute to wound debridement by removal of dead tissue. However, proteinase activity may also affect host tissue remodelling processes. Objectives

To identify proteolytic enzymes derived from L. sericata ES products with activities against fibrin and extracellular matrix (ECM) components. Methods

Larval proteinase activities were assayed in vitro using class-specific substrates and inhibitors. Their action against fibrin and ECM components was examined using sodium dodecyl sulphate–polyacrylamide gel electrophoresis. Results

Three classes of proteolytic enzyme were detected in the secretions using fluorescein isothiocyanate-labelled casein as a model substrate. The predominant activity belonged to serine proteinases (pH optima 8–9) of two different subclasses (trypsin-like and chymotrypsin-like), with a weaker aspartyl proteinase (pH 5) and a metalloproteinase (pH 9) with exopeptidase characteristics also present. Using skin-relevant ECM components as substrates L. sericata ES products solubilized fibrin clots and degraded fibronectin, laminin and acid-solubilized collagen types I and III. Hydrolysis of ECM macromolecules was inhibited by preincubating ES products with phenylmethylsulphonyl fluoride but not 4-amidinophenylmethylsulphonyl fluoride, indicating that degradation was due to the ‘chymotrypsin-like’ serine proteinase. Conclusions

These data suggest that a combination of L. sericata ES proteinases involving chymotrypsin-like and trypsin-like activities could potentially influence wound healing events when maggots are introduced into necrotic and infected wounds, with the chymotrypsin-like activity involved in the remodelling of ECM components.

Keywords: extracellular matrix components; chymotrypsin; serine proteinase; wound healing

Document Type: Research article

DOI: 10.1046/j.1365-2133.2003.04935.x

Affiliations: 1: Immune Modulation Group, School of Pharmaceutical Sciences, Boots Science Building, University of Nottingham, Nottingham NG7 2RD, U.K. 2: Schools of Environmental and Life Sciences and Continuing Education, University of Nottingham, Nottingham NG7 2RD, U.K. 3: Isis Innovation, University of Oxford, Ewert House, Summertown, Oxford OX2 7SG, U.K. 4: Royal Brompton Hospital, Sydney Street, London SW1 6NP, U.K. 5: Abney Court, Bourne End, Bucks SL8 5 DL, U.K.

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Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds