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Literature summary for 3.4.22.71 extracted from

  • 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.
    Structural and computational studies of the Staphylococcus aureus sortase B-substrate complex reveal a substrate-stabilized oxyanion hole (2014), J. Biol. Chem., 289, 8891-8902.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.22.71

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) pLysS Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment) 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

Protein Variants

Protein Variants Comment 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 UniProt Comment Textmining
Staphylococcus aureus A0A0H2XI76
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) pLysS by affinity chromatography Staphylococcus aureus

Synonyms

Synonyms Comment Organism
SrtB
-
 Staphylococcus aureus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Staphylococcus aureus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Staphylococcus aureus

General Information

General Information Comment 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