title: The metalloprotease of Listeria monocytogenes is activated by intramolecular autocatalysis.
abstract: The metalloprotease (Mpl) of Listeria monocytogenes is a thermolysin-like protease that mediates the maturation of a broad-range phospholipase C, whose function contributes to the ability of this food-borne bacterial pathogen to survive intracellularly. Mpl is made as a proprotein that undergoes maturation by proteolytic cleavage of a large N-terminal prodomain. In this study, we identified the N terminus of mature Mpl and generated Mpl catalytic mutants to investigate the mechanism of Mpl maturation. We observed that Mpl activity was a prerequisite for maturation, suggesting a mechanism of autocatalysis. Furthermore, using a strain of L. monocytogenes expressing both the wild-type form and a catalytic mutant form of Mpl simultaneously, we determined that in vivo maturation of Mpl occurs exclusively by an intramolecular autocatalysis mechanism.
authors.affiliation: Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Alan Pavinski
authors.initials: AP
authors.lastname: Bitar
authors.forename: Min
authors.initials: M
authors.lastname: Cao
authors.forename: Hélène
authors.initials: H
authors.lastname: Marquis
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism; D002384:Catalysis; D017931:DNA Primers; D004789:Enzyme Activation; D005838:Genotype; D007700:Kinetics; D008089:Listeria monocytogenes / Q000201:enzymology* / Q000235:genetics; D045726:Metalloproteases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D010446:Peptide Fragments; D011994:Recombinant Proteins; D010738:Type C Phospholipases; D045726:Metalloproteases