Information on EC 3.4.24.74 - fragilysin

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The expected taxonomic range for this enzyme is: Bacteroides fragilis

EC NUMBER
COMMENTARY
3.4.24.74
-
RECOMMENDED NAME
GeneOntology No.
fragilysin
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Broad proteolytic specificity, bonds hydrolysed including -Gly-/-Leu-, -Met-/-Leu-, -Phe-/-Leu-, -Cys-/-Leu-, Leu-/-Gly
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Baceroides fragilis enterotoxin
-
-
-
-
Baceroides fragilis enterotoxin
-
-
Baceroides fragilis enterotoxin
Bacteroides fragilis 086-5443-2-2
-
-
-
Bacteroides fragilis enterotoxin
-
-
Bacteroides fragilis toxin
-
-
-
-
Bacteroides fragilis toxin
O86049
-
Bacteroides fragilis toxin
Bacteroides fragilis 086-5443-2-2, Bacteroides fragilis 86-5443-2-2
-
-
-
Bacteroides fragilis toxin
Bacteroides fragilis ATCC 43858
O86049
-
-
Bacteroides fragilis toxin
Bacteroides fragilis I-1345
-
-
-
BFT
Bacteroides fragilis 086-5443-2-2, Bacteroides fragilis 86-5443-2-2, Bacteroides fragilis I-1345, Bacteroides fragilis NCTC 11295
-
-
-
BFT-3
O86049
isozyme
BFT-3
Bacteroides fragilis ATCC 43858
O86049
isozyme
-
Enterotoxin
-
-
-
-
fragilysin-3
O86049
isozyme
fragilysin-3
Bacteroides fragilis ATCC 43858
O86049
isozyme
-
Korea-BFT
O86049
-
additional information
O05091, O86049, Q9S5W0
the enzyme is a member of the matrix metalloprotease family of proteins
CAS REGISTRY NUMBER
COMMENTARY
188596-63-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
bft-1 is the most prevalent subtype, followed by bft-2 and a new bft subtype
-
-
Manually annotated by BRENDA team
enterotoxigenic strain ATCC 43858, and strains isolated from stool samples of calves with diarrhea containing gene bft
-
-
Manually annotated by BRENDA team
enterotoxigenic strains, isozyme Bft-1 encoded by gene bft-1
SwissProt
Manually annotated by BRENDA team
enterotoxigenic strains, isozyme Bft-2 encoded by gene bft-2
SwissProt
Manually annotated by BRENDA team
enterotoxigenic strains, isozyme Bft-3 encoded by gene bft-3
SwissProt
Manually annotated by BRENDA team
strain 086-5443-2-2
-
-
Manually annotated by BRENDA team
strain 086-5443-2-2, strain NCTC 9343
-
-
Manually annotated by BRENDA team
strain I-1345
-
-
Manually annotated by BRENDA team
strain NCTC 11295
-
-
Manually annotated by BRENDA team
the amount of produced enzyme depends on the composition of growth medium
-
-
Manually annotated by BRENDA team
two distinct isotypes of BFT: BFT-1 from strain VPI 13784 and BFT-2 from strain 86-5443-2-2
-
-
Manually annotated by BRENDA team
Bacteroides fragilis 086-5443-2-2
strain 086-5443-2-2
-
-
Manually annotated by BRENDA team
Bacteroides fragilis 86-5443-2-2
-
-
-
Manually annotated by BRENDA team
Bacteroides fragilis ATCC 43858
-
SwissProt
Manually annotated by BRENDA team
Bacteroides fragilis I-1345
strain I-1345
-
-
Manually annotated by BRENDA team
Bacteroides fragilis NCTC 11295
strain NCTC 11295
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
role of BFT in human host beta-defensin 2 induction from intestinal epithelial cells, HT-29 and Caco-2, requiring the presence of NF-kappaB binding sites. Also binding of NF-kappaB to the hBD-2 promoter is required for the induction of the hBD-2 gene in BFT-stimulated intestinal epithelial cells, while AP-1 is not involved, regulation, overview. Heat-labile enterotoxin BFT plays an essential role in host mucosal inflammation
physiological function
-
the enzyme plays an essential role in mucosal inflammation and affects the expression of ICAM-1 and monocytic adhesion to endothelial cells. Stimulation of HUVECs and rat aortic endothelial cells with Bacteroides fragilis enterotoxin results in the induction of ICAM-1 expression
physiological function
-
recombinant Bacteroides fragilis toxin (BFT, 5 nM) upregulates spermine oxidase in human colonic epithelial cell lines HT-29/c1 and T-84, resulting in spermine oxidase-dependent generation of reactive oxygen species and induction of gamma-H2A.x, a marker of DNA damage. BFT exposure did not increase spermidine/spermine-N1-acetyltransferase or N1-acetylpolyamine oxidase expression
physiological function
Bacteroides fragilis 86-5443-2-2
-
recombinant Bacteroides fragilis toxin (BFT, 5 nM) upregulates spermine oxidase in human colonic epithelial cell lines HT-29/c1 and T-84, resulting in spermine oxidase-dependent generation of reactive oxygen species and induction of gamma-H2A.x, a marker of DNA damage. BFT exposure did not increase spermidine/spermine-N1-acetyltransferase or N1-acetylpolyamine oxidase expression
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Abz-Asp-Tyr-Val-Ala-Ser-Glu-Lys(Dnp) + H2O
?
show the reaction diagram
-
-
-
-
?
Abz-Gly-Ile-Val-Arg-Ala-Lys(Dnp) + H2O
?
show the reaction diagram
-
-
-
-
?
Abz-Lys-Asp-Glu-Ser-Tyr-Arg-Lys(Dnp) + H2O
?
show the reaction diagram
-
-
-
-
?
Abz-Thr-Val-Leu-Glu-Arg-Ser-Lys(Dnp) + H2O
?
show the reaction diagram
-
-
-
-
?
Abz-Tyr-Gly-Lys-Arg-Val-Phe-Lys(Dnp) + H2O
?
show the reaction diagram
-
-
-
-
?
Abz-Val-Lys-Phe-Tyr-Asp-Ile-Lys(Dnp) + H2O
?
show the reaction diagram
-
-
-
-
?
Actin + H2O
Fragements of actin
show the reaction diagram
-
initial cleavage at Gly-Met, followed by Thr-Leu
-
-
actin + H2O
?
show the reaction diagram
-
-
-
-
?
actin + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
actin + H2O
?
show the reaction diagram
-
-
-
-
?
alpha1-proteinase inhibitor + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
?
show the reaction diagram
Bacteroides fragilis ATCC 43858
O86049
-
-
-
?
Azocoll + H2O
Fragements of azocoll
show the reaction diagram
-
-
-
-
azocollagen + H2O
?
show the reaction diagram
-
-
-
-
?
BODIPY FL-casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
Bacteroides fragilis ATCC 43858
O86049
-
-
-
?
collagen type IV + H2O
?
show the reaction diagram
-
-
-
-
?
E-cadherin + H2O
?
show the reaction diagram
-
-
-
-
?
E-cadherin + H2O
?
show the reaction diagram
O86049
-, cleavage of the zonula adherens protein
-
?
E-cadherin + H2O
?
show the reaction diagram
-
cleaves the extracellular domain
-
-
?
E-cadherin + H2O
?
show the reaction diagram
-
neither human nor rat E-cadherin are the cellular receptor for BFT
-
-
?
E-cadherin + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
the enzyme hydrolyzes the extracellular domain of several protein substrate, e.g. of E-cadherin to interrupt the intercellular adhesion increasing the permeability of the epithelium, and causing intracellular redistribution of actin with morphologic changes and release of beta-catenin
-
-
?
E-cadherin + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
the enzyme hydrolyzes the extracellular domain of the substrate
-
-
?
E-cadherin + H2O
?
show the reaction diagram
-
in vivo substrate
-
-
?
E-cadherin + H2O
?
show the reaction diagram
Bacteroides fragilis I-1345
-
neither human nor rat E-cadherin are the cellular receptor for BFT
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
-
-
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
-
-
-
-
?
Fibrinogen + H2O
?
show the reaction diagram
Bacteroides fragilis ATCC 43858
O86049
-
-
-
?
Fibrinogen + H2O
Fragements of fibrinogen
show the reaction diagram
-
hydrolysis to a lesser degree than other substrates
-
-
Fibronectin + H2O
?
show the reaction diagram
-
-
-
-
?
Fibronectin + H2O
?
show the reaction diagram
Bacteroides fragilis ATCC 43858
O86049
-
-
-
?
Fragilysin + H2O
Fragements of fragilysisn
show the reaction diagram
-
autodigestion
-
-
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Gelatin + H2O
Fragements of gelatin
show the reaction diagram
-
-
-
-
human complement C3 + H2O
?
show the reaction diagram
-
-
-
-
?
Mca-Arg-Pro-Lys-Pro-Val-Glu-Nva-Trp-Arg-Lys(Dnp)-NH2 + H2O
?
show the reaction diagram
-
-
-
-
?
myosin + H2O
?
show the reaction diagram
-
-
-
-
?
myosinO
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
myosinO + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
p-phenylazobenzyl-oxycarbonyl-Pro-Leu-Gly-Pro-D-Arg + H2O
?
show the reaction diagram
-
-
-
-
?
Proteins + H2O
?
show the reaction diagram
-
the enzyme increases the paracellular permeability of the intestinal epithelium by digestion of the intestinal light junctions and cell-to-cell contacts, it does not disrupt cell membranes
-
-
-
Proteins + H2O
?
show the reaction diagram
-
proteolytic action on the actin of the cytoskeleton
-
-
-
Proteins + H2O
?
show the reaction diagram
-
active in lamb, rabbit, rat ileum and colon, it causes the production of fluids and hemorrhage, it increases the levels of Na+ and Cl- in intestinal loops due to tissue damage
-
-
-
Tropomyosin + H2O
Fragements of tropomyosin
show the reaction diagram
-
hydrolysis to a lesser degree than other substrates
-
-
tropomyosin + H2O
?
show the reaction diagram
-
-
-
-
?
Mca-Pro-Leu-Gly-Leu-Dap(Dnp)-Ala-Arg-NH2 + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
cleaves at Cys-Leu, Ser-Leu, Thr-Leu, Gly-Leu or Leu-Gly peptide bonds in a number of proteins
-
-
?
additional information
?
-
O05091, O86049, Q9S5W0
the cytopathic effect of the enzyme is only seen in cells that can form tight junction and polarized monolayers, the enzyme affects the barrier function of the epithelium and decreases the transpithelial electrical resistance of monolayers, overview, the enzyme stimulates the recruitment of polymorphs to the lamina propria and seceretion of interleukin-8, epithelial neutrophil-activating peptide-78, and growth-related oncogene-alpha from colonocytes, overview
-
-
-
additional information
?
-
O05091, O86049, Q9S5W0
the cytopathic effect of the enzyme is only seen in cells that can form tight junction and polarized monolayers, the enzyme affects the barrier function of the epithelium and decreases the transpithelial electrical resistance of monolayers, overview, the enzyme stimulates the recruitment of polymorphs to the lamina propria and secretion of interleukin-8, epithelial neutrophil-activating peptide-78, and growth-related oncogene-alpha from colonocytes, overview
-
-
-
additional information
?
-
-
the enterotoxin has cytotoxic activity, however strains isolated from stool samples of calves with diarrhea contain the gene bft, but do not show cytotoxic activity against HT29 cells, overview
-
-
-
additional information
?
-
O05091, O86049, Q9S5W0
fragilysin is a zinc metalloprotease toxin with a C-terminal zinc-binding protease domain
-
-
-
additional information
?
-
-
isozyme fargilysin-3 does not cleave azoalbumin
-
-
-
additional information
?
-
Bacteroides fragilis ATCC 43858
O86049
isozyme fargilysin-3 does not cleave azoalbumin
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
actin + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
E-cadherin + H2O
?
show the reaction diagram
O86049
cleavage of the zonula adherens protein
-
?
E-cadherin + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
the enzyme hydrolyzes the extracellular domain of several protein substrate, e.g. of E-cadherin to interrupt the intercellular adhesion increasing the permeability of the epithelium, and causing intracellular redistribution of actin with morphologic changes and release of beta-catenin
-
-
?
E-cadherin + H2O
?
show the reaction diagram
-
in vivo substrate
-
-
?
myosinO
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
myosinO + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
Proteins + H2O
?
show the reaction diagram
-
the enzyme increases the paracellular permeability of the intestinal epithelium by digestion of the intestinal light junctions and cell-to-cell contacts, it does not disrupt cell membranes
-
-
-
Proteins + H2O
?
show the reaction diagram
-
proteolytic action on the actin of the cytoskeleton
-
-
-
Proteins + H2O
?
show the reaction diagram
-
active in lamb, rabbit, rat ileum and colon, it causes the production of fluids and hemorrhage, it increases the levels of Na+ and Cl- in intestinal loops due to tissue damage
-
-
-
Fibrinogen + H2O
?
show the reaction diagram
O05091, O86049, Q9S5W0
-
-
-
?
additional information
?
-
O05091, O86049, Q9S5W0
the cytopathic effect of the enzyme is only seen in cells that can form tight junction and polarized monolayers, the enzyme affects the barrier function of the epithelium and decreases the transpithelial electrical resistance of monolayers, overview, the enzyme stimulates the recruitment of polymorphs to the lamina propria and seceretion of interleukin-8, epithelial neutrophil-activating peptide-78, and growth-related oncogene-alpha from colonocytes, overview
-
-
-
additional information
?
-
O05091, O86049, Q9S5W0
the cytopathic effect of the enzyme is only seen in cells that can form tight junction and polarized monolayers, the enzyme affects the barrier function of the epithelium and decreases the transpithelial electrical resistance of monolayers, overview, the enzyme stimulates the recruitment of polymorphs to the lamina propria and secretion of interleukin-8, epithelial neutrophil-activating peptide-78, and growth-related oncogene-alpha from colonocytes, overview
-
-
-
additional information
?
-
-
the enterotoxin has cytotoxic activity, however strains isolated from stool samples of calves with diarrhea contain the gene bft, but do not show cytotoxic activity against HT29 cells, overview
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Zn2+
-
metalloprotease, 1 gatom of Zn2+ per mol of enzyme
Zn2+
-
stimulates at lower concentrations
Zn2+
O05091, O86049, Q9S5W0
zinc metalloprotease toxin with a C-terminal zinc-binding protease domain; zinc metalloprotease toxin with a C-terminal zinc-binding protease domain; zinc metalloprotease toxin with a C-terminal zinc-binding protease domain
Zn2+
-
dependent on
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2,2'-dipyridyl
-
-
5,7-dihydroxy-3',4',6-trimethoxyflavone
-
inhibits the BFT induced nuclear factor-kappaB activation, interleukin-8 secretion, cyclo-oxygenase-2 expression and activated I-kappa-B-alpha and I-kappa-B-kinase signals in human colon epithelial cell line HT-29. Analysis of the mechanism of inhibition
Alpha-macroglobulin
-
moderate inhibitor
-
diethyl dicarbonate
-
-
diethyldicarbonate
-
moderate inhibitor
Diethylenetriaminepentaacetic acid
-
-
dithiothreitol
-
-
genistein
-
broad spectrum tyrosine kinase inhibitor, significantly inhibits BFT-stimulated interleukin-8 production
germ agglutinin
-
inhibits BFT biological activity on HT29/C1 cells but not binding of Alexa 488-labeled BFT to HT29/C1 cells
-
PP2
-
selective Src-family tyrosine kinase inhibitor, significantly inhibits ca. 73% of BFT-stimulated interleukin-8 production
SB203580
-
p38 inhibitor, 0.01 mM inhibits ca. 80% of BFT-stimulated interleukin-8 production
subtilisin A
-
inhibits both BFT biological activity on HT29/C1 cells and nearly 90% of binding of Alexa 488-labeled BFT
-
Thermolysin
-
inhibits both BFT biological activity on HT29/C1 cells and nearly 90% of binding of Alexa 488-labeled BFT
-
thimerosal
-
i.e. sodium ethylmercurithiosalicylate
tyrphostin AG1478
-
selective epidermal growth factor receptor tyrosine kinase inhibitor, significantly inhibits ca. 64% of BFT-stimulated interleukin-8 production
U126
-
inhibits both basal and BFT-augmented extracellular signal-related kinase phosphorylation, 0.01 mM inhibits ca. 80% of BFT-stimulated interleukin-8 production
Zn2+
-
10 mM, slightly inhibitory
Zn2+
-
inhibits at higher concentrations
Zn2+
-
activity is significantly impaired by excess zinc
GM6001
-
significantly reduces Alexa 488-labeled BFT binding
additional information
-
not inhibitory are phenylmethylsulfonylfluoride, 4-(2-aminoethyl)-benzenesulfonylfluoride, tosylamide-2-phenylethyl chloromethylketone, tosyl-Lys chloromethylketone
-
additional information
-
not inhibited by lectins, sugars or mixed bovine brain gangliosides
-
additional information
-
activity is completely abolished by standard zinc-chelating agents
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
specific activity is expressed as a cytotoxic effect in cell-rounding titers of Ht-29 cells (i.e. colon carcinoma cells)
additional information
-
-
BFT stimulates apoptosis of human colon epithelial cell line HT-29. BFT suppresses apoptosis by increasing the cellular expression of cellular inhibitor of apoptosis protein-2 resulting in a delayed apoptosis
additional information
-
-
purified BFT does not directly stimulate lymphocyte proliferation, dendritic cell stimulation, or Toll-like receptor activation
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
-
O05091, O86049, Q9S5W0
-
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
the enterotoxin fragilysin is secreted from enterotoxic strains of Bacillus fragilis
-
Manually annotated by BRENDA team
-
the enzyme is secreted
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
18000
22000
-
gel fitration
20000
-
-
gel electrophoresis and Western blot analysis, mature processed BFT in all mutants
20000
-
-
Western blot analysis of cell-free culture supernatants of strain NCTC 9343 expressing mutants BFT-C2, BFT-C4 and BFT-A394E, and of whole cell lysates in preperations of strain NCTC 9343 expressing wild type BFT
20600
-
-
cleaved to its enzymatically active secreted form
44000
-
-
gel electrophoresis and Western blot analysis, unprocessed BFT in mutants H348D and M366R
44000
-
-
unprocessed enzyme, Western blot analysis of whole cell lysates in preperations of strain NCTC 9343 expressing wild type BFT
44400
-
-
preproprotein
additional information
-
-
analysis of BFT-2 by native PAGE reveals bands at 36000 Da, 90000 Da and 200000 Da. Both the 36000 Da and higher-molecular-mass bands are biologically active when tested in the HT29/C1 cell assay
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
O05091, O86049, Q9S5W0
x * 44000, preenzyme, x * 20000, mature enzyme; x * 44000, preenzyme, x * 20000, mature enzyme; x * 44000, preenzyme, x * 20000, mature enzyme
?
-
x * about 20000, SDS-PAGE
?
O86049
x * 21000, SDS-PAGE
?
Bacteroides fragilis ATCC 43858
-
x * 21000, SDS-PAGE
-
monomer
-
modelling of a three-dimensional structure
monomer
-
1 * 20600, SDS-PAGE
monomer
-
1 * 19000, SDS-PAGE
additional information
-
BFT-2 reveals a major band of 20000 Da with faint bands in some preparations at 70000 Da and 100000 Da, BFT-1 reveals a major band at 22000-23000 Da with fainter bands in some preparations at 70000 Da and 100000 Da, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sitting drop vapor diffusion method, using 100 mM sodium citrate dihydrate, 20% (w/v) PEG 3000, pH 5.5
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
O05091, O86049, Q9S5W0
stable over a wide range; stable over a wide range; stable over a wide range
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
-20
-
-
stable at
4
20
-
unstable and rapidly autodigests above 37°C
4
-
-
stable at
55
-
-
unstable above
additional information
-
O05091, O86049, Q9S5W0
stable over a wide range; stable over a wide range; stable over a wide range
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
BFT-1 and BFT-2 are partially cleaved by trypsin, 1:2 w/w, after 24 h. The pattern of tryptic peptides from BFT-1 and BFT-2 is identical
-
Stable to freeze-drying
-
stable to treatment with trypsin, chymotrypsin, and Dispase I
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
2000-3000fold purified by sequential ammonium sulfate precipitation, ion-exchange chromatography, hydrophobic chromatography, high resolution ion-exchange chromatography
-
BFT-1 and VFT-2
-
native enzyme from culture supernatant
-
nickel-affinity column chromatography
-
purified to homogeneity by urea-based purification method
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
analysis of the effect of enterotoxigenic Bacteroides fragilis in specific pathogen-free C57BL/6J mouse and germfree 129S6/SvEv mouse respectively
-
determination of pathogenicity islet
-
expressed in Escherichia coli Origami-2 (DE3) cells
-
expression of H348D in Escherichia coli
-
gene bft-1; gene bft-2; gene bft-3
O05091, O86049, Q9S5W0
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A394E
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mutation does not affect BFT processing or secretion
E349A
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expression of BFT similar to that of wild type
E349A
-
residual activity against fibrinogen and fluorophore-labeled casein
G355R
-
expression of BFT similar to that of wild type
H348D
-
smaller amounts of BFT compared to wild type
H352Y
-
expression of BFT similar to that of wild type
H352Y
-
Alexa-488-labeled BFT binding to HT29/C1 not blocked
H358Y
-
expression of BFT similar to that of wild type
E349A
Bacteroides fragilis ATCC 43858
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residual activity against fibrinogen and fluorophore-labeled casein
-
M366R
-
smaller amounts of BFT compared to wild type
additional information
-
BFT-C4, which has reduced BFT biological activity, shows a significant decrease in HT29/C1 cell binding
additional information
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C-terminal truncation mutants BFT-C2, BFT-C4, BFT-C7, BFT-C8, BFT-C9, BFT-C11, BFT-C18, deletion of only 2 amino acid residues at the C-terminus significantly reduces BFT biological activity and deletion of eight or more amino acid residues obliterated BFT biological activity, BFT mutants lacking seven or fewer amino acid residues in the C-terminal region are processed and expressed similar to wild-type BFT
H352Y
Bacteroides fragilis I-1345
-
Alexa-488-labeled BFT binding to HT29/C1 not blocked
-
additional information
Bacteroides fragilis I-1345
-
BFT-C4, which has reduced BFT biological activity, shows a significant decrease in HT29/C1 cell binding
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
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among stool strains from colon cancer patients, bft-1 is found to be more common than bft-2, isoform bft-1 is also found in almost all isolates from extraintestinal sites, strain NCTC 11295 harbours the isoform bft-2
medicine
Bacteroides fragilis NCTC 11295
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among stool strains from colon cancer patients, bft-1 is found to be more common than bft-2, isoform bft-1 is also found in almost all isolates from extraintestinal sites, strain NCTC 11295 harbours the isoform bft-2
-
medicine
-
stimulates interleukin-8 secretion by human intestinal epithelial HT29/C1 cells, biological active BFT, but not serum, is required for interleukin-8 production, NF-kB, tyrosine kinase and mitogen activated protein kinase signaling pathways are involved in BFT-induced interleukin-8 stimulation
additional information
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single point mutations in the zinc-binding metalloprotease motif do not affect BFT processing but do reduce or eliminate BFT biologic activity in vitro
additional information
-
intestinal epithelial cells in vitro possess a specific membrane BFT receptor that is distinct from E-cadherin
medicine
Bacteroides fragilis 086-5443-2-2
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stimulates interleukin-8 secretion by human intestinal epithelial HT29/C1 cells, biological active BFT, but not serum, is required for interleukin-8 production, NF-kB, tyrosine kinase and mitogen activated protein kinase signaling pathways are involved in BFT-induced interleukin-8 stimulation
-
additional information
Bacteroides fragilis I-1345
-
intestinal epithelial cells in vitro possess a specific membrane BFT receptor that is distinct from E-cadherin
-