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show all sequences of 3.4.22.71

Structural and computational studies of the Staphylococcus aureus sortase B-substrate complex reveal a substrate-stabilized oxyanion hole

Jacobitz, A.W.; Wereszczynski, J.; Yi, S.W.; Amer, B.R.; Huang, G.L.; Nguyen, A.V.; Sawaya, M.R.; Jung, M.E.; McCammon, J.A.; Clubb, R.T.; J. Biol. Chem. 289, 8891-8902 (2014)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) pLysS
Staphylococcus aureus
Crystallization (Commentary)
Crystallization
Organism
sortase B enzyme in a covalent complex with an analogue of its NPQTN sorting signal substrate, hanging drop vapour diffusion method, micing of 0.150 mMSrtB-NPQT in 10 mM Tris-HCl, pH 7.0, 20 mM NaCl, with reservoir solution containing 2.8 M ammonium sulfate, 70 mM sodium citrate, pH 5.0, X-ray diffraction structure determination and analysis at 2.49 A resolution, molecular replacement method
Staphylococcus aureus
Engineering
Amino acid exchange
Commentary
Organism
C223A
site-directed mutagenesis, inactive mutant
Staphylococcus aureus
D225A
site-directed mutagenesis, the mutant exhibits nonspecific proteolytic activity
Staphylococcus aureus
H130A
site-directed mutagenesis, inactive mutant
Staphylococcus aureus
R233A
site-directed mutagenesis, inactive mutant
Staphylococcus aureus
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Staphylococcus aureus
Q2FHU6
-
-
Purification (Commentary)
Commentary
Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) pLysS by affinity chromatography
Staphylococcus aureus
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Staphylococcus aureus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Staphylococcus aureus
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) pLysS
Staphylococcus aureus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
sortase B enzyme in a covalent complex with an analogue of its NPQTN sorting signal substrate, hanging drop vapour diffusion method, micing of 0.150 mMSrtB-NPQT in 10 mM Tris-HCl, pH 7.0, 20 mM NaCl, with reservoir solution containing 2.8 M ammonium sulfate, 70 mM sodium citrate, pH 5.0, X-ray diffraction structure determination and analysis at 2.49 A resolution, molecular replacement method
Staphylococcus aureus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C223A
site-directed mutagenesis, inactive mutant
Staphylococcus aureus
D225A
site-directed mutagenesis, the mutant exhibits nonspecific proteolytic activity
Staphylococcus aureus
H130A
site-directed mutagenesis, inactive mutant
Staphylococcus aureus
R233A
site-directed mutagenesis, inactive mutant
Staphylococcus aureus
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) pLysS by affinity chromatography
Staphylococcus aureus
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Staphylococcus aureus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Staphylococcus aureus
General Information
General Information
Commentary
Organism
evolution
the enzyme is a member of the sortase cysteine transpeptidase family. Members of this enzyme superfamily are widely distributed in Gram-positive bacteria that frequently utilize multiple sortases to elaborate their peptidoglycan, and the family members have a conserved active site His-Cys-Arg triad that joins a sorting signal located at the C-terminus of their protein substrate to an amino nucleophile located on the cell surface
Staphylococcus aureus
additional information
analysis of the substrate binding structure of the enzyme using SrtB-NPQT complexes, by computational modeling, molecular dynamics simulations, and targeted amino acid mutagenesis revealing that the backbone amide of Glu224 and the side chain of Arg233 form an oxyanion hole in sortase B that stabilizes high energy tetrahedral catalytic intermediates. A highly conserved threonine residue within the bound sorting signal substrate facilitates construction of the oxyanion hole by stabilizing the position of the active site arginine residue via hydrogen bonding
Staphylococcus aureus
physiological function
sortase cysteine transpeptidases covalently attach proteins to the bacterial cell wall or assemble fiber-like pili that promote bacterial adhesion
Staphylococcus aureus
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme is a member of the sortase cysteine transpeptidase family. Members of this enzyme superfamily are widely distributed in Gram-positive bacteria that frequently utilize multiple sortases to elaborate their peptidoglycan, and the family members have a conserved active site His-Cys-Arg triad that joins a sorting signal located at the C-terminus of their protein substrate to an amino nucleophile located on the cell surface
Staphylococcus aureus
additional information
analysis of the substrate binding structure of the enzyme using SrtB-NPQT complexes, by computational modeling, molecular dynamics simulations, and targeted amino acid mutagenesis revealing that the backbone amide of Glu224 and the side chain of Arg233 form an oxyanion hole in sortase B that stabilizes high energy tetrahedral catalytic intermediates. A highly conserved threonine residue within the bound sorting signal substrate facilitates construction of the oxyanion hole by stabilizing the position of the active site arginine residue via hydrogen bonding
Staphylococcus aureus
physiological function
sortase cysteine transpeptidases covalently attach proteins to the bacterial cell wall or assemble fiber-like pili that promote bacterial adhesion
Staphylococcus aureus
Other publictions for EC 3.4.22.71
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
732174
Jacobitz
Structural and computational s ...
Staphylococcus aureus
J. Biol. Chem.
289
8891-8902
2014
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3
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718136
Spirig
Sortase enzymes in Gram-positi ...
Staphylococcus aureus
Mol. Microbiol.
82
1044-1059
2011
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718269
Kang
Crystal structure of Spy0129, ...
Streptococcus pyogenes, Streptococcus pyogenes M1 SF370
PLoS ONE
6
e15969
2011
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Oh
Therapeutic effect of (Z)-3-(2 ...
Staphylococcus aureus, Staphylococcus aureus Newman
Biochem. Biophys. Res. Commun.
396
440-444
2010
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693248
Mariscotti
The Listeria monocytogenes sor ...
Listeria monocytogenes
J. Biol. Chem.
284
6140-6146
2009
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680796
Maresso
Activation of inhibitors by so ...
Bacillus anthracis
J. Biol. Chem.
282
23129-23139
2007
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680881
Bentley
Engineering the substrate spec ...
Staphylococcus aureus
J. Biol. Chem.
282
6571-6581
2007
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663737
Oh
Inhibition of sortase-mediated ...
Staphylococcus aureus
Appl. Microbiol. Biotechnol.
70
102-106
2006
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664280
Kang
Flavonols inhibit sortases and ...
Staphylococcus aureus
Biol. Pharm. Bull.
29
1751-1755
2006
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680473
Maresso
Surface protein IsdC and sorta ...
Bacillus anthracis
J. Bacteriol.
188
8145-8152
2006
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665174
Zink
Importance of srtA and srtB fo ...
Bacillus anthracis
Infect. Immun.
73
5222-5228
2005
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665617
Marraffini
Anchor structure of staphyloco ...
Staphylococcus aureus
J. Biol. Chem.
280
16263-16271
2005
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666393
Newton
The svpA-srtB locus of Listeri ...
Listeria monocytogenes
Mol. Microbiol.
55
927-940
2005
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666890
Pucciarelli
Identification of substrates o ...
Listeria monocytogenes
Proteomics
5
4808-4817
2005
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647690
Zong
The structure of sortase B, a ...
Staphylococcus aureus
Structure
12
105-112
2004
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647691
Bierne
Sortase B, a new class of sort ...
Listeria monocytogenes
J. Bacteriol.
186
1972-1982
2004
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666932
Zhang
Structures of sortase B from S ...
Bacillus anthracis, Staphylococcus aureus
Structure
12
1147-1156
2004
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647386
Jonsson
The role of Staphylococcus aur ...
Staphylococcus aureus
Microbes Infect.
5
775-780
2003
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