Proteolytic enzymes from Trichinella spiralis larvae.
Trichinella spiralis larvae infect their hosts by the penetration of small intestine enterocytes. The exact mechanism of penetration is unknown, but the presence of proteolytic enzymes is suspected. In this study, whole worm extracts and excretory-secretory (ES) components were obtained and their proteolytic enzymes examined. Enzymes from worm extracts were capable of hydrolysing azocoll, a general protease substrate in a wide range of pH (2-8), with maximal activity at pH 5. Trichinella spiralis larval enzymes were sensitive to metalloprotease and serine protease inhibitors. Three proteases were identified in worm extracts at molecular weight (MW) 48, 54 and 62 kDa by incorporating a gelatine substrate into a standard or a modified sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) set-up, in which we used low SDS concentration in gel and electrophoresis buffer (0.01%). Intact larvae incubated in a medium containing azocoll showed azocollytic activity. Subsequent analysis of ES products by modified SDS-PAGE in gels containing gelatine demonstrated the presence of three protease of apparent MW 33, 62 and 230 kDa.
Criado-Fornelio
A
A
Parasitology Laboratory, Faculty of Pharmacy, University of Alcalá de Henares, Madrid, Spain.
de Armas-Serra
C
C
Giménez-Pardo
C
C
Casado-Escribano
N
N
Jiménez-Gónzalez
A
A
Rodríguez-Caabeiro
F
F
D000818:Animals; D001391:Azo Compounds / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D003124:Colorimetry; D004396:Coloring Agents / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007814:Larva / Q000201:enzymology; D008970:Molecular Weight; D017160:Trichinella spiralis / Q000201:enzymology*
D001391:Azo Compounds; D004396:Coloring Agents; C028470:Azocoll; D003094:Collagen; D010450:Endopeptidases