Information on EC 3.4.22.B63 - sortase C

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.B63
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
sortase C
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
The enzyme from Bacillus anthracis cleaves the LPNT-/-A sorting signal of BasH and BasI, thereby anchoring both polypeptides to the cell wall envelope of sporulating bacilli [17074072]. The enzymes from Streptococcus pneumoniae and Enterococcus faecalis are required for efficient pilus assembly.
show the reaction diagram
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
BasH polypeptide + H2O
?
show the reaction diagram
BasI polypeptide + H2O
?
show the reaction diagram
Dabcyl-ancillary protein 1-Edans + H2O
?
show the reaction diagram
Dabcyl-ancillary protein 2-Edans + H2O
?
show the reaction diagram
Dabcyl-BP-1 peptide-Edans + H2O
?
show the reaction diagram
Dabcyl-KKVTIPQTGGIGT-EDANS + H2O
?
show the reaction diagram
Q3DMP3
-
-
-
?
surface protein containing a QVPTGV motif + H2O
?
show the reaction diagram
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SrtC2 anchors a surface protein containing a QVPTGV motif (followed by a hydrophobic region and a charged tail) to the cell wall
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-
?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
BasH polypeptide + H2O
?
show the reaction diagram
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sortase C cleaves the LPNTA sorting signal of BasH and BasI, thereby anchoring both polypeptides to the cell wall envelope of sporulating bacilli. Sortase C specifically recognizes and cleaves the LPNTA motif. SrtCDELTAN (with a replacement of the N-terminal signal peptide of sortase C for six histidy) cleaves peptides encompassing the LPNTA motif between threonine and alanine. Sortase C substrate BasH is expressed in the forespore
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-
?
BasI polypeptide + H2O
?
show the reaction diagram
Dabcyl-ancillary protein 1-Edans + H2O
?
show the reaction diagram
Dabcyl-ancillary protein 2-Edans + H2O
?
show the reaction diagram
Dabcyl-BP-1 peptide-Edans + H2O
?
show the reaction diagram
surface protein containing a QVPTGV motif + H2O
?
show the reaction diagram
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SrtC2 anchors a surface protein containing a QVPTGV motif (followed by a hydrophobic region and a charged tail) to the cell wall
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-
?
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Q3DMP3
SrtC1 is not involved in calcium binding
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-(trimethylammonium)ethyl thiol
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methylmethane-thiosulphonate
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hypothesis that the thiol of Cys181 functions as the active site SrtCDELTAN
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00358 - 0.06036
Dabcyl-ancillary protein 1-Edans
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0.01421 - 0.02733
Dabcyl-ancillary protein 2-Edans
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0.02156 - 0.04385
Dabcyl-BP-1 peptide-Edans
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0.0138 - 0.0294
Dabcyl-KKVTIPQTGGIGT-EDANS
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00104 - 0.0049
Dabcyl-ancillary protein 1-Edans
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0.00041 - 0.00113
Dabcyl-ancillary protein 2-Edans
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0.000731 - 0.00298
Dabcyl-BP-1 peptide-Edans
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16.2 - 810
Dabcyl-ancillary protein 1-Edans
202479
159 - 797
Dabcyl-ancillary protein 2-Edans
202480
243 - 922
Dabcyl-BP-1 peptide-Edans
202478
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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srtC is expressed during sporulation
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24010
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electrospray mass spectrometry analysis
28000
Q3DMP3
x * 28000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
Q3DMP3
x * 28000, SDS-PAGE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
apoprotein, hanging drop vapor diffusion method, using 15-25% (w/v) polyethylene glycol 3350 and 0.2 M Tris buffer (pH 8.0-8.8) and 0.2 M ammonium acetate at 4C. In complex with inhibitor 2-(trimethylammonium)ethyl thiol, hanging drop vapor diffusion method, using 1.6 M ammonium sulfate, 5% (w/v) polyethylene glycol 3350, and sodium cacodylate (pH 6.5) at 4C
hanging drop vapor diffusion method, using 10% (w/v) PEG monomethyl ether 2000 and 100 mM MES (pH 6.4)
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sitting drop vapor diffusion method, using 100 mM Tris (pH 8.5), 200 mM NaOAc, and 30% (w/v) PEG-4000
Q3DMP3
sitting drop vapor diffusion method
Q97SB8
three different crystals are obtained using both the sitting-drop and hanging-drop vapour-diffusion methods. The three crystals belonged to different space groups and diffracted to resolutions ranging between 2.3 and 1.7 A. One crystal form belongs to space group P212121, with unit cell parameters a = 48.9, b = 96.9, c = 98.9 A, alpha = beta = gamma = 90. The other two crystal forms belong to space group P222, with unit-cell parameters a = 48.8, b = 97.2, c= 99.2 A, alpha = beta = gamma = 90 and a = 48.6, b= 96.5, c = 98.8 A, alpha = beta = gamma = 90, respectively
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isofofm SrtC1, hanging drop vapor diffusion method, using 0.4 M sodium formate, 0.1 M Bis-Tris propane pH 6.5, and 22% (w/v) PEG 3350. Isoform SrtC1, hanging drop vapor diffusion method, using 0.26 M CaCl2, 19% (w/v) PEG 6000, 0.1 M HEPES pH 7.0
hanging drop vapor diffusion method, using 0.1 M Tris-HCl pH 8.5, 0.54 M sodium citrate, 8% (v/v) iso-propanol, 8% (v/v) tert-butanol
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
His-Trap column chromatography, and Superdex 75 gel filtration
Ni-NTA column chromatography
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Ni-NTA resin column chromatography, and Superdex 75 gel filtration
SrtCDELTAN (with a replacement of the N-terminal signal peptide of sortase C for six histidy) is expressed in Escherichia coli
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expressed in Escherichia coli B834(DE3) cells
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3) pLysS
Q97SB8
expressed in Escherichia coli M15 [pREP4] cells
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expressed in Escherichia coli Rosetta (DE3) pLysS cells
SrtC protein is cloned into plasmid pET24c and expressed in Escherichia coli without any tag
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SrtCDELTAN (with a replacement of the N-terminal signal peptide of sortase C for six histidy) is expressed in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C219A/H157A/R228A
Q3DMP3
inactive
D84A
Q3DMP3
the mutation in the lid does not abrogate pilus protein polymerization
Y86A
Q3DMP3
the mutant show an increased Km value compared to the wild type enzyme, the mutation in the lid does not abrogate pilus protein polymerization
F86A
the mutant shows decreased catalytic efficiency compared to the wild type
Y92A
the mutant shows increased catalytic efficiency compared to the wild type
F86A
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the mutant shows decreased catalytic efficiency compared to the wild type
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Y92A
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the mutant shows increased catalytic efficiency compared to the wild type
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additional information
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mutation or deletion of the positively charged domain flanking a transmembrane helix in SrtC abolishes both its retention at single foci and its function in efficient pilus assembly. This positively charged domain can act as a localization retention signal for the focal compartmentalization of membrane proteins