title: Synthesis and cleavage- secretion of enzymatically active rabbit angiotensin-converting enzyme in Pichia pastoris.
abstract: Many biologically important ectoproteins that are anchored in the plasma membrane via a hydrophobic domain undergo a proteolytic cleavage process, which releases the ectodomain to the extracellular milieu in a regulated fashion. Angiotensin-converting enzyme (ACE) is one such protein that is secreted from human and mouse cells by its cleavage at one of two alternative sites in the ectodomain. Here, we report similar cleavage-secretion of ACE in the yeast Pichia pastoris. The cleavage site used in yeasts was identical to one of the two sites used in mouse cells. Moreover, as in mammalian cells, ACE secretion in yeast was inhibited by compound 3, a potent inhibitor of the metzincin family of metalloproteases. ACE proteins cleavage-secreted from yeast and from mammalian cells had identical enzymatic properties. These results demonstrate the existence of a secretase activity in yeast whose properties closely resemble those of the mammalian ACE secretase.
authors.affiliation: Department of Molecular Cardiology, Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
authors.forename: R
authors.initials: R
authors.lastname: Sadhukhan
authors.forename: G C
authors.initials: GC
authors.lastname: Sen
authors.forename: I
authors.initials: I
authors.lastname: Sen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004274:DNA, Recombinant / Q000235:genetics; D010450:Endopeptidases / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D007703:Peptidyl-Dipeptidase A / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000201:enzymology / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D013737:Testis / Q000201:enzymology
chemical_list: D004274:DNA, Recombinant; D011994:Recombinant Proteins; D010450:Endopeptidases; D007703:Peptidyl-Dipeptidase A; C077675:angiotensin converting enzyme secretase