title: Characterization of the promastigote surface protease of Leishmania as a membrane-bound zinc endopeptidase.
abstract: The effects of a variety of inhibitors suggested that the promastigote surface protease (PSP) of Leishmania might be a zinc metalloprotease. To investigate this possibility, we conducted atomic emission and absorption spectroscopic analyses, which show that PSP contains 1 atom of zinc per 63-kDa monomer. Further studies showed that the enzyme can be biosynthetically labeled with 65ZnCl2. The comparison of the amino acid sequence of Leishmania major PSP with nine other zinc metalloproteinases revealed significant similarity in the area of their zinc-binding sites. These data show clearly that the promastigote surface protease of Leishmania is a zinc metalloproteinase. Secondary structure analysis by circular dichroism spectroscopy indicates that PSP contains over 40% beta-strand and less than 20% alpha-helical structure. The molecular masses of amphiphilic PSP (152 kDa) and of hydrophilic PSP (142 kDa), determined by quantitative electron scattering, suggest that the purified enzyme occurs in solution, and presumably at the cell surface, as a non-covalent homodimer.
authors.affiliation: Institut de Biochimie, Université de Lausanne, Epalinges, Switzerland.
authors.forename: J
authors.initials: J
authors.lastname: Bouvier
authors.forename: C
authors.initials: C
authors.lastname: Bordier
authors.forename: H
authors.initials: H
authors.lastname: Vogel
authors.forename: R
authors.initials: R
authors.lastname: Reichelt
authors.forename: R
authors.initials: R
authors.lastname: Etges
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002712:Chlorides / Q000378:metabolism; D002942:Circular Dichroism; D006801:Humans; D007895:Leishmania tropica / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000032:analysis / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012689:Sequence Homology, Nucleic Acid; D013054:Spectrophotometry, Atomic; D015032:Zinc / Q000032:analysis* / Q000378:metabolism; D017967:Zinc Compounds
chemical_list: D002712:Chlorides; D017967:Zinc Compounds; C016837:zinc chloride; D008666:Metalloendopeptidases; D015032:Zinc