The atypical amino-terminal LPNTG-containing domain of the pneumococcal human IgA1-specific protease is required for proper enzyme localization and function.
Streptococcus pneumoniae produces a zinc metalloproteinase, Iga, which cleaves human immunoglobulin A1 (IgA1), and whose activity is predominantly localized to the bacterial surface. However, proper surface localization is not predicted using current models, as the LPNTG sorting motif is located atypically near the amino- rather than the carboxy-terminus. The cell-associated form of Iga was confirmed to be external to the bacterial membrane, and while bound tightly, its attachment to the cell wall is non-covalent, but dependent on both a complete LPNTG sequence and sortase activity. Disruption of the region between the signal peptidase cleavage site and the LPNTG domain resulted in a localization defect, premature degradation, and an alteration of the ability of the enzyme to act on a monoclonal human IgA1 substrate and to enhance bacterial adherence, linking localization to enzyme function. Edman sequencing of cell-associated Iga determined that the enzyme is processed at an unexpected site downstream of the sorting signal yet still associates with the bacterial surface. Our results indicate a non-covalent re-association between the carboxy-terminal enzymatic domain and the cleaved, sorted amino-terminal localization domain. This amino-terminal motif is shared among the other zinc metalloproteinases in streptococci and suggests a novel conserved mechanism for the surface localization of protease activity.
Bender
Matthew H
MH
Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
Weiser
Jeffrey N
JN
D000595:Amino Acid Sequence; D019881:Aminoacyltransferases / Q000378:metabolism; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D002478:Cells, Cultured / Q000382:microbiology; D003546:Cysteine Endopeptidases / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D010457:Peptidoglycan / Q000378:metabolism; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D013296:Streptococcus pneumoniae / Q000201:enzymology* / Q000502:physiology
D001426:Bacterial Proteins; D010457:Peptidoglycan; D021382:Protein Sorting Signals; D019881:Aminoacyltransferases; C120542:sortase A; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase; D003546:Cysteine Endopeptidases