title: Surface acid proteinase (gp63) of Leishmania mexicana. A metalloenzyme capable of protecting liposome-encapsulated proteins from phagolysosomal degradation by macrophages.
abstract: Acid proteinase activity is associated with the major surface glycoprotein (gp63) of both extracellular promastigotes and intracellular amastigotes of the parasitic protozoan, Leishmania mexicana. The enzyme purified by monoclonal affinity chromatography from promastigotes is strongly inhibited by metal ion chelators, which is reversible by the addition of Zn(II). This proteinase loses its activity after dialysis against 1,10-phenanthroline. The apoenzyme thus prepared is reactivated substantially by Zn(II) and partially by Cu(II), Cd(II), Co(II), or Ni(II). From the recently published structure of the gene encoding gp63, we identify hitherto unrecognized sequences, which can be aligned to the consensus zinc-binding sites of other known metalloproteinases. Anti-gp63 polyclonal antibodies, but not the monoclonals, precipitate similar molecules from amastigotes. These molecules differ slightly from gp63 in electrophoretic mobility but have similar endopeptidase activity. Phagolysosomal degradation by macrophages of proteins entrapped in liposomes is prevented by coating them with native gp63. This protection is lost with heat denaturation of gp63 to kill its enzymatic activity. The proteolytic activity of the metalloenzyme on the surface of these parasites may thus protect their membrane from cytolytic damages during their survival, differentiation, and multiplication in the phagolysosomes of macrophages.
authors.affiliation: Department of Microbiology/Immunology, University of Health Sciences/Chicago Medical School.
authors.forename: G
authors.initials: G
authors.lastname: Chaudhuri
authors.forename: M
authors.initials: M
authors.lastname: Chaudhuri
authors.forename: A
authors.initials: A
authors.lastname: Pan
authors.forename: K P
authors.initials: KP
authors.lastname: Chang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D002614:Chelating Agents / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007894:Leishmania mexicana / Q000166:cytology / Q000201:enzymology*; D008247:Lysosomes / Q000469:parasitology; D008264:Macrophages / Q000469:parasitology / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010588:Phagosomes / Q000469:parasitology
chemical_list: D002614:Chelating Agents; D008562:Membrane Glycoproteins; D008666:Metalloendopeptidases