A novel extracellular calcium-dependent cysteine proteinase from Crithidia deanei.
An extracellular cysteine proteinase from an aposymbiotic strain of Crithidia deanei was purified 39-fold by a combination of anion-exchange and gel filtration chromatographies. The native molecular mass of this proteinase was estimated to be 225 kDa by gel filtration chromatography and it migrates in SDS-PAGE as a single band of 80 kDa. The optimal enzymatic activity on gelatin was found to occur in the presence of calcium at a neutral pH and at 28 degrees C. The enzyme was completely blocked by E-64 and EGTA, and partially inhibited by iodoacetamide, leupeptin, and EDTA. Compounds such as PMSF, aprotinin, and pepstatin weakly inhibited the enzyme. The protein purified in the present work shares some features with those of the family of neutral calcium-dependent cysteine proteinases named calpains, previously detected in the family Trypanosomatidae as cell-associated enzymes in Leishmania donovani and Trypanosoma brucei. The cysteine proteinase from C. deanei is distinct from the well-characterized mammalian calpains, but some degree of similarity is displayed to invertebrate calpain-related enzymes.
d'Avila-Levy
Claudia M
CM
Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, CCS, Bl I, 21941-590, Rio de Janeiro, RJ, Brazil.
Souza
Rodrigo F
RF
Gomes
Rosana C
RC
Vermelho
Alane B
AB
Branquinha
Marta H
MH
D000818:Animals; D002474:Cell-Free System; D002478:Cells, Cultured; D003421:Crithidia / Q000737:chemistry* / Q000201:enzymology*; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry / Q000378:metabolism; D004789:Enzyme Activation; D004795:Enzyme Stability; D005110:Extracellular Space / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D013045:Species Specificity; D013696:Temperature
D015853:Cysteine Proteinase Inhibitors; D003546:Cysteine Endopeptidases