Enzymatic Activity of the Staphylococcus aureus SplB Serine Protease is Induced by Substrates Containing the Sequence Trp-Glu-Leu-Gln

Proteases are of significant importance for the virulence of Staphylococcus aureus. Nevertheless, their subset, the serine protease-like proteins, remains poorly characterized. Here presented is an investigation of SplB protease catalytic activity revealing that the enzyme possesses exquisite specif...

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Veröffentlicht in:Journal of molecular biology 2008-05, Vol.379 (2), p.343-356
Hauptverfasser: Dubin, Grzegorz, Stec-Niemczyk, Justyna, Kisielewska, Magdalena, Pustelny, Katarzyna, Popowicz, Grzegorz M., Bista, Michal, Kantyka, Tomasz, Boulware, Kevin T., Stennicke, Henning R., Czarna, Anna, Phopaisarn, Mullika, Daugherty, Patrick S., Thøgersen, Ida B., Enghild, Jan J., Thornberry, Nancy, Dubin, Adam, Potempa, Jan
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container_end_page 356
container_issue 2
container_start_page 343
container_title Journal of molecular biology
container_volume 379
creator Dubin, Grzegorz
Stec-Niemczyk, Justyna
Kisielewska, Magdalena
Pustelny, Katarzyna
Popowicz, Grzegorz M.
Bista, Michal
Kantyka, Tomasz
Boulware, Kevin T.
Stennicke, Henning R.
Czarna, Anna
Phopaisarn, Mullika
Daugherty, Patrick S.
Thøgersen, Ida B.
Enghild, Jan J.
Thornberry, Nancy
Dubin, Adam
Potempa, Jan
description Proteases are of significant importance for the virulence of Staphylococcus aureus. Nevertheless, their subset, the serine protease-like proteins, remains poorly characterized. Here presented is an investigation of SplB protease catalytic activity revealing that the enzyme possesses exquisite specificity and only cleaves efficiently after the sequence Trp-Glu-Leu-Gln. To understand the molecular basis for such selectivity, we solved the three-dimensional structure of SplB to 1.8 Å. Modeling of substrate binding to the protease demonstrated that selectivity relies in part on a canonical specificity pockets-based mechanism. Significantly, the conformation of residues that ordinarily form the oxyanion hole, an essential structural element of the catalytic machinery of serine proteases, is not canonical in the SplB structure. We postulate that within SplB, the oxyanion hole is only formed upon docking of a substrate containing the consensus sequence motif. It is suggested that this unusual activation mechanism is used in parallel with classical determinants to further limit enzyme specificity. Finally, to guide future development, we attempt to point at likely physiological substrates and thus the role of SplB in staphylococcal physiology.
doi_str_mv 10.1016/j.jmb.2008.03.059
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subjects Amino Acid Sequence
Animals
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Binding Sites
Crystallography, X-Ray
Humans
Models, Molecular
Molecular Sequence Data
Molecular Structure
Peptide Library
Peptides - chemistry
Peptides - genetics
Peptides - metabolism
protease
Serine Endopeptidases - chemistry
Serine Endopeptidases - genetics
Serine Endopeptidases - metabolism
Staphylococcus aureus
Staphylococcus aureus - enzymology
structure
Substrate Specificity
title Enzymatic Activity of the Staphylococcus aureus SplB Serine Protease is Induced by Substrates Containing the Sequence Trp-Glu-Leu-Gln
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