A non-catalytic histidine residue influences the function of the metalloprotease of Listeria monocytogenes.

Mpl, a thermolysin-like metalloprotease, and PC-PLC, a phospholipase C, are synthesized as proenzymes by the intracellular bacterial pathogen Listeria monocytogenes. During intracellular growth, L. monocytogenes is temporarily confined in a membrane-bound vacuole whose acidification leads to Mpl autolysis and Mpl-mediated cleavage of the PC-PLC N-terminal propeptide. Mpl maturation also leads to the secretion of both Mpl and PC-PLC across the bacterial cell wall. Previously, we identified negatively charged and uncharged amino acid residues within the N terminus of the PC-PLC propeptide that influence the ability of Mpl to mediate the maturation of PC-PLC, suggesting that these residues promote the interaction of the PC-PLC propeptide with Mpl. In the present study, we identified a non-catalytic histidine residue (H226) that influences Mpl secretion across the cell wall and its ability to process PC-PLC. Our results suggest that a positive charge at position 226 is required for Mpl functions other than autolysis. Based on the charge requirement at this position, we hypothesize that this residue contributes to the interaction of Mpl with the PC-PLC propeptide.

[1]  Brian M. Forster,et al.  Protein transport across the cell wall of monoderm Gram‐positive bacteria , 2012, Molecular microbiology.

[2]  Brian M. Forster,et al.  The Metalloprotease of Listeria monocytogenes Is Regulated by pH , 2011, Journal of bacteriology.

[3]  H. Marquis,et al.  Differentiation of propeptide residues regulating the compartmentalization, maturation and activity of the broad-range phospholipase C of Listeria monocytogenes. , 2010, The Biochemical journal.

[4]  L. Nicholson,et al.  Elucidation of a pH-folding switch in the Pseudomonas syringae effector protein AvrPto , 2009, Proceedings of the National Academy of Sciences.

[5]  K. Stiasny,et al.  Identification of specific histidines as pH sensors in flavivirus membrane fusion , 2008, The Journal of cell biology.

[6]  M. Cao,et al.  The Metalloprotease of Listeria monocytogenes Is Activated by Intramolecular Autocatalysis , 2007, Journal of bacteriology.

[7]  J. Srivastava,et al.  Intracellular pH sensors: design principles and functional significance. , 2007, Physiology.

[8]  G. Thomas,et al.  Identification of a pH Sensor in the Furin Propeptide That Regulates Enzyme Activation* , 2006, Journal of Biological Chemistry.

[9]  Torsten Schwede,et al.  BIOINFORMATICS Bioinformatics Advance Access published November 12, 2005 The SWISS-MODEL Workspace: A web-based environment for protein structure homology modelling , 2022 .

[10]  R. Tweten,et al.  Molecular basis of listeriolysin O pH dependence. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Sergio A Hassan,et al.  Long dynamics simulations of proteins using atomistic force fields and a continuum representation of solvent effects: Calculation of structural and dynamic properties , 2005, Proteins.

[12]  H. Marquis,et al.  The Metalloprotease of Listeria monocytogenes Controls Cell Wall Translocation of the Broad-Range Phospholipase C , 2005, Journal of bacteriology.

[13]  Piero Fariselli,et al.  ConSeq: the identification of functionally and structurally important residues in protein sequences , 2004, Bioinform..

[14]  H. Marquis,et al.  Restricted Translocation across the Cell Wall Regulates Secretion of the Broad-Range Phospholipase C of Listeria monocytogenes , 2003, Journal of bacteriology.

[15]  Manuel C. Peitsch,et al.  SWISS-MODEL: an automated protein homology-modeling server , 2003, Nucleic Acids Res..

[16]  H. Bouwer,et al.  Isolation of Listeria monocytogenes mutants with high‐level in vitro expression of host cytosol‐induced gene products , 2003, Molecular microbiology.

[17]  H. Marquis,et al.  pH‐regulated activation and release of a bacteria‐associated phospholipase C during intracellular infection by Listeria monocytogenes , 2000, Molecular microbiology.

[18]  M. Loessner,et al.  Modified Listeria bacteriophage lysin genes (ply) allow efficient overexpression and one-step purification of biochemically active fusion proteins , 1996, Applied and environmental microbiology.

[19]  D. Portnoy,et al.  The two distinct phospholipases C of Listeria monocytogenes have overlapping roles in escape from a vacuole and cell-to-cell spread , 1995, Infection and immunity.

[20]  D. Portnoy,et al.  Dual roles of plcA in Listeria monocytogenes pathogenesis , 1993, Molecular microbiology.

[21]  P. Youngman,et al.  Use of a new integrational vector to investigate compartment-specific expression of the Bacillus subtilis spoIIM gene. , 1992, Biochimie.

[22]  P. Berche,et al.  Reduced virulence of a Listeria monocytogenes phospholipase-deficient mutant obtained by transposon insertion into the zinc metalloprotease gene , 1992, Infection and immunity.

[23]  P. Cossart,et al.  Identification of a new operon involved in Listeria monocytogenes virulence: its first gene encodes a protein homologous to bacterial metalloproteases , 1991, Infection and immunity.

[24]  D. Portnoy,et al.  Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes , 1989, The Journal of cell biology.

[25]  S. Ho,et al.  Site-directed mutagenesis by overlap extension using the polymerase chain reaction. , 1989, Gene.

[26]  S. Miyoshi,et al.  Microbial metalloproteases and pathogenesis. , 2000, Microbes and infection.

[27]  N. Guex,et al.  SWISS‐MODEL and the Swiss‐Pdb Viewer: An environment for comparative protein modeling , 1997, Electrophoresis.