The pathway of secretion of proteinases in Trichomonas vaginalis.
Trichomonads secrete large amounts of hydrolytic enzymes into liquid growth medium. Proteinase release by Trichomonas vaginalis has been quantified after resuspension of the parasite in a simple buffered maltose medium. After 6 h incubation, 70-90% of each of two cysteine proteinase activities, one towards benzyloxycarbonyl-arginyl-arginine 4-nitroanilide (Z-RR-Nan) and the other active towards N-benzoyl-prolyl-phenylalanyl-arginine 4-nitroanilide (Bz-PFR-Nan), was extracellular. This release was insensitive to changes in pH within the range from 5.5 to 8.6 but was partially inhibited by chloride ions. The secretion of activity towards Bz-PFR-Nan was temperature-sensitive but was still detectable down to 14 degrees C. Neither this nor other cysteine proteinase activities were detectable on the surface of parasites. Release was stimulated by various amines and monensin, suggesting that secretion was from or via acidic compartments. The intracellular activity towards Bz-PFR-Nan could be totally and irreversibly inhibited by treating the parasites with benzyloxycarbonyl-phenylalanyl-alanine diazomethylketone (Z-FA-DMK), without otherwise harming the cells. Regeneration and routing of the proteinases responsible for this activity was followed after removal of the inhibitor. There was a significant rise in the intracellular level of activity before it became detectable in the medium. The release of this activity was accelerated by amines and monensin, but the build-up of enzyme activity within cells was not prevented. Organelles containing cysteine proteinases banded as a single peak in Percoll density gradients. The density of these increased when cells were treated with dextran. The activity towards Bz-PFR-Nan which reappeared after Z-FA-DMK treatment has a similar distribution. The proteinase-containing fraction could be distinguished from an early (5 min) endosome fraction, suggesting that it was composed of late endosomes/lysosomes. Thus these results imply that the secretion pathway for proteinases necessarily involves lysosomes/late endosomes.
Scott
D A
DA
Department of Zoology, University of Glasgow, Scotland, U.K.
North
M J
MJ
Coombs
G H
GH
D000588:Amines / Q000494:pharmacology; D000818:Animals; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000378:metabolism; D007700:Kinetics; D008247:Lysosomes / Q000201:enzymology*; D008985:Monensin / Q000494:pharmacology; D013696:Temperature; D014246:Trichomonas vaginalis / Q000201:enzymology*
D000588:Amines; D008985:Monensin; D006867:Hydrolases; D003546:Cysteine Endopeptidases