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PDBsum entry 4ink
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PDB id:
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Hydrolase
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Title:
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Crystal structure of spld protease from staphylococcus aureus at 1.56 a resolution
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Structure:
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Serine protease spld. Chain: a. Fragment: unp residues 37-239. Engineered: yes
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Source:
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Staphylococcus aureus subsp. Aureus. Organism_taxid: 93061. Strain: nctc 8325. Gene: saouhsc_01938, spld. Expressed in: escherichia coli. Expression_system_taxid: 562
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Resolution:
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1.56Å
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R-factor:
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0.180
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R-free:
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0.210
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Authors:
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M.Zdzalik,M.Kalinska,P.Cichon,M.Wysocka,J.Stec-Niemczyk, H.R.Stennicke,A.Jabaiah,M.Markiewicz,B.Wladyka,P.S.Daugherty, A.Lesner,K.Rolka,A.Dubin,J.Potempa,G.Dubin
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Key ref:
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M.Zdzalik
et al.
(2013).
Biochemical and structural characterization of SplD protease from Staphylococcus aureus.
Plos One,
8,
e76812.
PubMed id:
DOI:
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Date:
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04-Jan-13
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Release date:
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30-Oct-13
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PROCHECK
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Headers
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References
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Q2FXC5
(SPLD_STAA8) -
Serine protease SplD from Staphylococcus aureus (strain NCTC 8325 / PS 47)
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Seq:
Struc:
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239 a.a.
203 a.a.
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Key: |
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PfamA domain |
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Secondary structure |
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CATH domain |
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DOI no:
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Plos One
8:e76812
(2013)
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PubMed id:
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Biochemical and structural characterization of SplD protease from Staphylococcus aureus.
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M.Zdzalik,
M.Kalinska,
M.Wysocka,
J.Stec-Niemczyk,
P.Cichon,
N.Stach,
N.Gruba,
H.R.Stennicke,
A.Jabaiah,
M.Markiewicz,
S.Kedracka-Krok,
B.Wladyka,
P.S.Daugherty,
A.Lesner,
K.Rolka,
A.Dubin,
J.Potempa,
G.Dubin.
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ABSTRACT
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Staphylococcus aureus is a dangerous human pathogen. A number of the proteins
secreted by this bacterium are implicated in its virulence, but many of the
components of its secretome are poorly characterized. Strains of S. aureus can
produce up to six homologous extracellular serine proteases grouped in a single
spl operon. Although the SplA, SplB, and SplC proteases have been thoroughly
characterized, the properties of the other three enzymes have not yet been
investigated. Here, we describe the biochemical and structural characteristics
of the SplD protease. The active enzyme was produced in an Escherichia coli
recombinant system and purified to homogeneity. P1 substrate specificity was
determined using a combinatorial library of synthetic peptide substrates showing
exclusive preference for threonine, serine, leucine, isoleucine, alanine, and
valine. To further determine the specificity of SplD, we used high-throughput
synthetic peptide and cell surface protein display methods. The results not only
confirmed SplD preference for a P1 residue, but also provided insight into the
specificity of individual primed- and non-primed substrate-binding subsites. The
analyses revealed a surprisingly narrow specificity of the protease, which
recognized five consecutive residues (P4-P3-P2-P1-P1') with a consensus motif of
R-(Y/W)-(P/L)-(T/L/I/V)↓S. To understand the molecular basis of the strict
substrate specificity, we crystallized the enzyme in two different conditions,
and refined the structures at resolutions of 1.56 Å and 2.1 Å. Molecular
modeling and mutagenesis studies allowed us to define a consensus model of
substrate binding, and illustrated the molecular mechanism of protease
specificity.
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