title: Sequential autolytic processing activates the zymogen of Arg-gingipain.
abstract: Most proteases are synthesized as inactive precursors to protect the synthetic machinery of the cell and allow timing of activation. The mechanisms used to render latency are varied but tend to be conserved within protease families. Proteases belonging to the caspase family have a unique mechanism mediated by transitions of two surface loops, and on the basis of conservation of mechanism one would expect this to be preserved by caspase relatives. We have been able to express the full-length precursor of the Arg-specific caspase relative from the bacterium Porphyromonas gingivalis, Arg-gingipain-B, and we show that it contains N- and C-terminal extensions that render a low amount of latency, meaning that the zymogen is substantially active. Three sequential autolytic processing steps at the N and C terminus are required for full activity, and the N-propeptide may serve as an intramolecular chaperone rather than an inhibitory peptide. Each step in activation requires the previous step, and an affinity probe reveals that incremental activity enhancements are achieved in a stepwise manner.
authors.affiliation: Burnham Institute, La Jolla, California 92037, USA.
authors.forename: Jowita
authors.initials: J
authors.lastname: Mikolajczyk
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Boatright
authors.forename: Henning R
authors.initials: HR
authors.lastname: Stennicke
authors.forename: Tamim
authors.initials: T
authors.lastname: Nazif
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
authors.forename: Guy S
authors.initials: GS
authors.lastname: Salvesen
mesh_terms: D018829:Adhesins, Bacterial; D002384:Catalysis; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins / Q000378:metabolism*; D016966:Porphyromonas gingivalis / Q000201:enzymology*; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D018829:Adhesins, Bacterial; D004792:Enzyme Precursors; D000080867:Gingipain Cysteine Endopeptidases; D006388:Hemagglutinins; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases