An early event in the Trypanosoma cruzi cell invasion process, the recruitment of host lysosomes, led us to investigate the involvement of signal transduction. Infective trypomastigotes were found to contain a soluble Ca2+-signaling activity for mammalian cells that is sensitive to protease inhibitors. Inhibitor and substrate utilization profiles were used to purify a candidate peptidase for involvement in this process, from which we isolated a full-length cDNA clone. The sequence revealed a novel enzyme, denominated T. cruzi oligopeptidase B, which is homologous to members of the prolyl oligopeptidase family of serine hydrolases, known to participate in the maturation of biologically active peptides. The T. cruzi oligopeptidase B was expressed as a fully active product in Escherichia coli, and antibodies to the recombinant enzyme inhibited both peptidase activity and Ca2+ signaling induced in normal rat kidney cells by trypomastigote extracts. Our data suggest that the T. cruzi oligopeptidase B participates in processing events in the cytoplasm of the parasites, generating a factor with Ca2+-signaling activity for mammalian cells.
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10 February 1997
Article|
February 10 1997
A Cytosolic Serine Endopeptidase from Trypanosoma cruzi Is Required for the Generation of Ca2+ Signaling in Mammalian Cells
Barbara A. Burleigh,
Barbara A. Burleigh
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
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Elisabet V. Caler,
Elisabet V. Caler
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
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Paul Webster,
Paul Webster
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
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Norma W. Andrews
Norma W. Andrews
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
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Barbara A. Burleigh
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
Elisabet V. Caler
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
Paul Webster
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
Norma W. Andrews
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002
This work was funded by National Institutes of Health grant R01AI32056 to N.W. Andrews. B. Burleigh was supported by a postdoctoral fellowship from the Medical Research Council of Canada.
Address all correspondence to Norma W. Andrews, Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520-8002. Tel.: (203) 785-4314. Fax: (203) 785-7226. E-mail: [email protected]
Received:
September 17 1996
Revision Received:
November 13 1996
Online ISSN: 1540-8140
Print ISSN: 0021-9525
1997
J Cell Biol (1997) 136 (3): 609–620.
Article history
Received:
September 17 1996
Revision Received:
November 13 1996
Citation
Barbara A. Burleigh, Elisabet V. Caler, Paul Webster, Norma W. Andrews; A Cytosolic Serine Endopeptidase from Trypanosoma cruzi Is Required for the Generation of Ca2+ Signaling in Mammalian Cells. J Cell Biol 10 February 1997; 136 (3): 609–620. doi: https://doi.org/10.1083/jcb.136.3.609
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