Information on EC 3.6.3.50 - protein-secreting ATPase

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The expected taxonomic range for this enzyme is: Bacteria, Archaea

EC NUMBER
COMMENTARY hide
3.6.3.50
-
RECOMMENDED NAME
GeneOntology No.
protein-secreting ATPase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + H2O = ADP + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase (protein-secreting)
A non-phosphorylated, non-ABC (ATP-binding cassette) ATPase that is involved in protein transport. There are several families of enzymes included here, e.g. ATP-hydrolysing enzymes of the general secretory pathway (Sec or Type II), of the virulence-related secretory pathway (Type III) and of the conjugal DNA-protein transfer pathway (Type IV). Type II enzymes occur in bacteria, archaea and eucarya, whereas type III and type IV enzymes occur in bacteria where they form components of a multi-subunit complex.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-83-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
type III secretion system
-
-
Manually annotated by BRENDA team
type IV secretion system
-
-
Manually annotated by BRENDA team
gene bsaS
UniProt
Manually annotated by BRENDA team
strain CWL029, ATCC VR1310
-
-
Manually annotated by BRENDA team
type III secretion system; type II secretion system
-
-
Manually annotated by BRENDA team
type II secretion system
-
-
Manually annotated by BRENDA team
-
Swissprot
Manually annotated by BRENDA team
strain DK1622
-
-
Manually annotated by BRENDA team
strain DK1622
-
-
Manually annotated by BRENDA team
pv. gysophila; type III secretion system
-
-
Manually annotated by BRENDA team
type III secretion system
-
-
Manually annotated by BRENDA team
subsp. carotovora, type II secretion apparatus
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain PAK, genes pilB, pilT and pilU
-
-
Manually annotated by BRENDA team
type III secretion system
-
-
Manually annotated by BRENDA team
type III secretion system
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
Pathovar citri
-
-
Manually annotated by BRENDA team
pv. vesicatoria
-
-
Manually annotated by BRENDA team
pv. oryzae
-
-
Manually annotated by BRENDA team
type III secretion system
-
-
Manually annotated by BRENDA team
Yersinia enterolytica
-
-
-
Manually annotated by BRENDA team
type III secretion system YscN ATPase
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + H2O
ADP + phosphate
show the reaction diagram
HopB1/in + ATP + H2O
HopB1/out + ADP + phosphate
show the reaction diagram
-
intrinsic protein substrate, type II effector of pathogen
-
-
?
HopPtoN/in + ATP + H2O
HopPtoN/out + ADP + phosphate
show the reaction diagram
-
Hrp outer protein effector of pathogen, that is translocated into host cells in enzyme-dependent secretion
-
-
?
HrpJ/in + ATP + H2O
HrpJ/out + ADP + phosphate
show the reaction diagram
-
intrinsic protein substrate, its secretion is required for pathogenicity and translocation of effectors into plant cells
-
-
?
HrpK/in + ATP + H2O
HrpK/out + ADP + phosphate
show the reaction diagram
-
intrinsic protein substrate, C-terminal half of protein is required for translocation
-
-
?
MgATP + H2O
MgADP + phosphate
show the reaction diagram
-
-
-
-
?
YopR/in + ATP + H2O
YopR/out + ADP + phosphate
show the reaction diagram
-
eleven N-terminal amino acids of YopR sectretion substrate function as secretion signal required for binding to enzyme
-
-
?
additional information
?
-
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
ATP + H2O
ADP + phosphate
show the reaction diagram
HopB1/in + ATP + H2O
HopB1/out + ADP + phosphate
show the reaction diagram
-
intrinsic protein substrate, type II effector of pathogen
-
-
?
HopPtoN/in + ATP + H2O
HopPtoN/out + ADP + phosphate
show the reaction diagram
-
Hrp outer protein effector of pathogen, that is translocated into host cells in enzyme-dependent secretion
-
-
?
HrpJ/in + ATP + H2O
HrpJ/out + ADP + phosphate
show the reaction diagram
-
intrinsic protein substrate, its secretion is required for pathogenicity and translocation of effectors into plant cells
-
-
?
HrpK/in + ATP + H2O
HrpK/out + ADP + phosphate
show the reaction diagram
-
intrinsic protein substrate, C-terminal half of protein is required for translocation
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
stimulation of sectretion
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,3-dicyclohexylcarbodiimide
-
0.01 mM, 76% residual activity
5'-adenylylimidodiphosphate
-
1 mM, 12% residual activity
ADP
-
competitive inhibitor
ATPase inhibitor compound 939
-
-
FliH
Because FliH suppresses ATP hydrolysis by FliI, FliH coordinates ATP hydrolysis by FliI with protein export. The first 20 residues of FliI (FliIEN) not only regulate FliI ring formation but also are involved in the interaction with FliH
-
KNO3
-
10 mM, 74% residual activity
N'-[(E)-(2,4-dihydroxyphenyl)methylidene]-4-nitrobenzohydrazide
A0A0C6EWK5
a type III secretion system inhibitor
N'-[(E)-(3,5-dibromo-2-hydroxyphenyl)methylidene]pyridine-3-carbohydrazide
A0A0C6EWK5
a type III secretion system inhibitor
NaCl
about 78% reduced activity at 500 mM NaCl
Sodium azide
-
10 mM, 10% residual activity
YsaL
identification and evaluation of a negative regulator of YsaN, a hypothetical protein YE3555, termed YsaL. Purified YsaL is dimeric in solution and strongly associates with YsaN to form a stable heterotrimeric YsaL-YsaN complex with a stoichiometry of 2:1. The N terminal 6-20 residues of YsaN are invariably required for stable YsaL-YsaN complex formation. YsaL inhibits the ATPase activity of YsaN with a maximum inhibition at the molar ratio 2:1 (YsaL: YsaN). Protein YE3555 has a close evolutionary relationship with the T3S ATPase regulators. Identification of critical residues of YsaN for stable YsaL-YsaN complex formation, overview
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YscN
-
intrinsic regulatory protein, noncompetitive inhibition by allosteric binding
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cardiolipin
FliH
-
the stimulation of the FliI ATPase activity by FliJ occurs either via a transient FliJ-FliI interaction or via interactions of FliJ with other flagellar components, e.g. the FliH protein, the flagellar counterpart of HrpE, with a predicted coiled-coil structure. FliH is a known regulator of FliI activity that binds to FliI and suppresses its oligomerization and ATPase activity
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phosphatidylglycerol
-
via the EpsE/cyto-EpsL complex
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0726 - 1.3
ATP
0.22
MgATP
-
pH 7.5, 37°C, ATPase activity of N-terminal cytoplasmic domain
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.052 - 447
Abz-VAA
-
0.00062 - 0.35
ATP
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.06
ADP
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00108
-
recombinant PilT mutant H229A
0.00275
-
recombinant PilT mutant E163Q
0.0029
-
recombinant His6-tagged PilB
0.005
-
recombinant wild-type PilU
0.0074
-
recombinant wild-type PilT
0.0108
-
recombinant wild-type PilB
0.15
Q9Z748
pH 7.0, 37°C
0.36
purified recombinant SsaN-Myc-His6 enzyme, pH 8.0, 37°C
0.55
-
wild-type full-length CdsN and C-terminal truncation mutant of CdsN, ATPase activity
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8
-
ATPase activity of N-terminal cytoplasmic domain
additional information
-
secretion of the avirulence proteins HrmA and AvrPto from Pseudomonas syringae pathovars is highest at pH 6.0 of the culture medium
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 42
-
ATPase activity of N-terminal cytoplasmic domain
additional information
-
secretion of the avirulence proteins HrmA and AvrPto from Pseudomonas syringae pathovars is highest at growth temperature 18-22°C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 42
-
-
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.1
-
recombinant GST-tagged enzyme, sequence calculation
8
Q9Z748
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
a number of the Out apparatus components possess domains in the cytoplasm and/or the periplasm with potential for protein-protein interactions which facilitate the secretion of periplasmic enzyme intermediates across the outer membrane to the external milieu. OutC is a cytoplasmic membrane protein with a single transmembrane domain and a large hydrophilic periplasmic domain, OutF is a cytoplasmic membrane protein with three transmembrane domains, a small periplasmic loop, a large cytoplasmic loop and an N-terminal cytoplasmic domain
-
Manually annotated by BRENDA team
additional information
assembly of the ATPase complex to the export gate, model, overview
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
39000
-
1 * 62000, recombinant His-tagged PilB, SDS-PAGE, x * 63800, about, His6-tagged PilB, sequence calculation, 1 * 39000, recombinant His-tagged PilT, SDS-PAGE, 1 * 41000, about, His6-tagged PilT, sequence calculation
40000
-
PilT, gel filtration
41000
-
1 * 62000, recombinant His-tagged PilB, SDS-PAGE, x * 63800, about, His6-tagged PilB, sequence calculation, 1 * 39000, recombinant His-tagged PilT, SDS-PAGE, 1 * 41000, about, His6-tagged PilT, sequence calculation
45000
-
determined by SDS-PAGE, HisEscN
47500
Q9Z748
x * 47500, calculated
48100
-
x * 48100, about, sequence calculation, SDS-PAGE, CdsN forms oligomers and high-molecular-weight multimers, possibly dodecamers
49500
12 * 49500, recombinant His-tagged enzyme, SDS-PAGE. The enzyme exists in solution as higher order oligomer. The ATPase activity of oligomeric YsaN is several fold higher than the activity of the monomeric form
51800
-
deduced molecular mass
56000
x * 56000
63800
-
1 * 62000, recombinant His-tagged PilB, SDS-PAGE, x * 63800, about, His6-tagged PilB, sequence calculation, 1 * 39000, recombinant His-tagged PilT, SDS-PAGE, 1 * 41000, about, His6-tagged PilT, sequence calculation
64000
-
x * 64000, YscN, SDS-PAGE
83000
3 * 83000, GspE-cyto-GspL complex, crystal structure analysis
250000
GspE-cyto-GspL complex trimer, crystal structure analysis
290000
-
gel filtration
300000
sedimentation analysis
327000
-
hexamer, determined by blue native electrophoresis
350000
-
about
575000
sedimentation analysis, predominant form
603000
recombinant His-tagged enzyme dodecamer, gel filtration
622000
-
dodecamer, determined by blue native electrophoresis
3500000
sedimentation analysis
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dodecamer
hexamer
homohexamer
monomer
oligomer
trimer
3 * 83000, GspE-cyto-GspL complex, crystal structure analysis
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
lipoprotein
-
the lipidated protein MxiM, required in type III secretion, is anchored to the inner leaflet of the outer membrane
proteolytic modification
-
EscC, 56000 Da preprotein is cleaved to 54000 mature protein, EscV, 75000 Da preprotein with sec-dependent signal sequence, 72000 mature protein, EscN, 49000 Da cytoplasmic protein with no signal sequence
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
structure analysis
crystals of selenomethionine-substituted afGspE are grown in the presence of 10 mM AMP-PNP and 10 mM Mg2+, a resolution of 2.95 A is attained
-
the crystal structure of the catalytic domain of the prototypical T3SS ATPase EscN of Escherichia coli is presented at a resolution of 1.8 A
-
cryo-electron microscopy, enzyme dodecamer comprises two hexameric rings that are stacked face-to-face by the association of their C-terminal regions
-
purified recombinant wild-type enzyme, hanging drop vapor diffusion method, mixing of 0.002 ml of 1.7 mg/ml protein in 20 mM Tris, pH 7.5, 0.1 M potassium chloride, and 0.01 M tris(2-carboxyethyl)phosphine with 0.001 ml of crystallization solution containing 0.5 M ammonium sulfate, 0.1 M sodium citrate tribasic dihydrate, pH 5.6, and 10% v/v Jeffamine M-600, and 200 nl of 0.1 M L-proline, the drops are initially dehydrated against 0.5 ml of 1.5 M ammonium sulfate, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement and modeling
major cytoplasmic domain of EpsL protein that is part of the ATPase complex
-
periplasmic domain of EpsM protein, fold is a circular permutation of the ferredoxin fold
-
recombinant enzyme GspE-Hcp1 fusion protein hexamers, sitting drop vapour diffusion method, mixing of 0.001 ml protein solution containing 20 mM Tris-HCl, pH 8.0, 500 mM NaCl, 5% v/v glycerol, 1 mM TCEP/HCl, 5 mM AMPPNP, and 5 mM MgCl2 with different precipitant solutions, containing for DN1GspEEpsE-GSGSGS-Hcp1 7% PEG 6000 and 0.1 M bicine, pH 9.0, for DN1GspEEpsE-KLASGAGH-Hcp1 16% PEG 300, 0.2 M ammonium sulfate, 0.1 M Bis Tris, pH 6.1, 5 mM ADP, 5 mM MgCl2, 5 mM AlCl3, and 15 mM NaF, for DELTAN1GspEEpsE-KLASGAG-Hcp1 7% PEG 3350, 0.12 M ammonium citrate pH 7.0, 5 mM ADP, and 5 mM MgCl2, and for DELTAN1GspEEpsE-KLASG-Hcp1 12.5% PEG 20000, 0.1 M bicine pH 9.0, and 2% v/v 1,4-dioxane, at 4°C, X-ray diffraction structure determination and analysis at 4.09-7.6 A resolution
purified recombinant enzyme GspE in complex with cyto-GspL, sitting drop vapor diffusion method, by mixing of 0.001 ml of protein in 20 mM HEPES, pH7.5, 200 mM NaCl buffer with 0.001 ml of reservoir solution containing 0.2 M Na malonate pH 7.0, 18% PEG 3350, at 21°C, X-ray diffraction structure determination and analysis at 2.83 A resolution, molecular replacement using the structures of Vibrio cholerae DELTAN1EGspE (PDB ID 1P9R) and the Vibrio cholerae N1E-cyto-GspL complex (PDB ID 2BH1) as search models
21 kDa N-terminal domain of enzyme in two conformational states
-
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
stability of HrcN depends on the conserved HrcL protein, which interacts with HrcN in vitro and in vivo
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by affinity chromatography, using Talon, Co2+, resin and glutathione-Sepharose beads, and by gel filtration
-
EpsE is copurified with the cytoplasmic domain of EpsL to test whether the ATPase activity of EpsE can be modulated by EpsL, complexes are purified using metal affinity chromatography and gel filtration, monomeric EpsE is obtained following thrombin cleavage of a GST-EpsE fusion protein
-
His-tagged EscN is purified from the soluble fraction using nickel-chelating Sepharose, the tag is cleaved with thrombin, cleaved product is purified by Mono-Q anion exchange
-
N-terminal cytoplasmic domain, with His-tag
-
partially by subcellular fractionation of wild-type strain DK1622 expressing GFP-tagged wild-type PilB and PilT, recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography
-
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
recombinant decahistidinyl derivative
recombinant FLAG-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by affinity chromatography
recombinant GST-tagged full-length CdsN and CdsN C-terminal truncation mutant from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography
-
recombinant GST-tagged YscP by glutathione affinity chromatography, YscP-GST co-purifies with YscN, YscL, YscQ, and YscO
-
recombinant His10-FLAG3-tagged InvC/FLAG3-tagged OrgB complexes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
recombinant SsaN-Myc-His6, EscN-Myc-His6, and SsaNR192G-Myc-His6 wefrom Escherichia coli by nickel affinity chromatography
recombinant Strep-tagged wild-type and mutant enzymes by streptavidin affinity chromatography
-
separation of the recombinant Ysc proteins
-
soluble part of recombinant N-terminally His-tagged wild-type enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration, recombinant His-tagged truncation mutant enzymes from Escherichia coli strain BL21(DE3) are unfolded, dialysed and refolded and purified by nickel affinity chromatography and gel filtration, recombinant untagged wild-type enzyme
using a Ni-NTA agarose column
-
XpsE-Strep and its variants are purified from the Strep-Tactin Sepharose column, the XpsE with His6-tag is purified by eluting from nickel nitrilotriacetate resin
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Yersinia enterocolitica LJ4036, a strain in which the wild-type copy of yscP is deleted from the virulence plasmid
-
expression in Escherichia coli
expression of C-terminally c-Myc-tagged ATPase HrcN under control of the lac promoter into Xanthomonas campestris strains 85-10DhrcQ and 85*DhrcQ, co-expression with GST-HrcQ and C-terminally c-Myc-tagged HrcL in Escherichia coli
-
expression of N-terminally His-tagged wild-type and deletion mutant Flil enzymes, and expression of the deletion mutants in Salmonella typhimurium strain SJW1103
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expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain JM109, expression of GFP-tagged wild-type PilB and PilT in strain DK1622
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expression of YscN and other protein complex components in Escherichia coli
-
expresssion of GST-tagged full-length CdsN and a C-terminal truncation mutant of CdsN in Escherichia coli strain BL21(DE3)
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gene hrcN, cloned from strain 85-10, co-expression with c-Myc-tagged hrcL in Escherichia coli strain BL21(DE3), expression of Strep-tagged HrcN and Strep-tagged HrcN mutant G175C
-
gene invC, recombinant expression of His10-FLAG3-tagged InvC/FLAG3-tagged OrgB complexes in Escherichia coli strain BL21(DE3)
gene pscN, phylogenetic analysis and tree
A0A0C6EWK5
gene ssaN, recombinant expression of wild-type and mutant enzymes tagged with hemagglutinin, c-Myc, a FLAG- or a His6-tag, in Escherichia coli and Salmonella enterica, co-expression with SsaK-2HA and SsaQ-2HA fusion proteins
gene xpsE, recombinant expression of C-terminally ECFP-tagged enzyme in the xpsE-null strain XC1723 complementing the strain and restoring the secretion activity
gene xpsE, xpsE gene regulation and promoter activity analysis, overview
-
gene ysaN, recombinant expression of N-terminally His-tagged wild-type and of His-tagged truncation mutant enzymes in Escherichia coli strain BL21(DE3), expression of untagged wild-type enzyme
gene yscP, expression of GST-fusion YscP in wild-type strain W22703 and strain EC2 DELTA(yscM1 yscM2)
-
genes pilB, pilT and pilU from strain PAK, expression of wild-type and mutant enzymes in Escherichia coli strain Bl21(DE3). Expression of YFP-PilT, YFP-PilB, and YFP-PilU fusion enzymes, which retain their characteristic pattern of polar localization, with the exception of the Walker A mutant of YFP-PilT
-
into the pET19b vector for expression in Escherichia coli BL21DE3 pLysS cells
-
into the pET28a expression vector for expression in Escherichia coli BL21DE3 cells
-
nine pcfF expression plasmids are constructed, different in the combination of the promotor, Pnis, P23, Ptac, PT7, the affinity tag, His6, GST, and the cloned PcfC fragment, complete cDNA, deltaN103 or mutant K156T
-
plasmids pEpsE/EpsL(1-253)His6 and pEpsE/EpsL(1-242)His6 using pET21d+ are constructed, for expression of monomeric EpsE the vector GST-EpsE is constructed
-
recombinant decahistidinyl derivative
recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
recombinant expression of FLAG-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
recombinant expression of the enzyme GspE as fusion proteins with Pseudomonas aeruginosa Hcp1, i.e. DELTAN1GspEEpsE-KLASGHcp1, DELTAN1GspEEpsE-GSGSGS-Hcp1, DELTAN1GspEEpsE-KLASGAGHcp1, and DELTAN1GspEEpsE-KLASGAGH-Hcp1 showing homogeneous hexamer formation, several variants of DN1GspEEpsE-Hcp1 fusions with different linker sequences are constructed
the cloned Erwinia chrysanthemi Hrp type III protein secretion system functions in Escherichia coli to deliver Pseudomonas syringae Avr signals to plant cells and to secrete Avr proteins in culture
-
using a pET29a based plasmid and pET15b for overexpressing of the protein in Escherichia coli BL21DE3 pLysS cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
deltaN103
-
pcfF mutant lacking codon 1 to 103
K156T
-
pcfF mutant
EscNDELTA102
-
EscN mutant
EscNDELTA102-V393P
-
mutant, used for crystallization
EscNDELTA7
-
EscN mutant
EscNDELTA7-R366D
-
EscN mutant
EscNDELTA8
-
EscN mutant
G182C
-
EscN mutant
K183E
-
EscN mutant
R207G
-
EscN mutant
R209H
-
EscN mutant
R240H
-
EscN mutant
R366D
-
EscN mutant
R113E
-
no ATPase activity
R133E
-
dramatic decrease in ATPase activity
R18A
-
dramatic decrease in hexameric particles, increase ATPase activity
E205A
-
a Walker B box substitution in PilT, leads to strong reduction of ATPase activity due to direct interference with ATP hydrolysis
E391A
-
a Walker B box substitution in PilB, leads to strong reduction of ATPase activity due to direct interference with ATP hydrolysis
K137A
-
a Walker A box substitution in PilT, leads to abolishment of ATPase activity due to indirect interference withe ATP binding
K327A
-
a Walker A box substitution in PilB, leads to strong reduction of ATPase activity due to indirect interference withe ATP binding
E205A
-
a Walker B box substitution in PilT, leads to strong reduction of ATPase activity due to direct interference with ATP hydrolysis
-
E391A
-
a Walker B box substitution in PilB, leads to strong reduction of ATPase activity due to direct interference with ATP hydrolysis
-
K137A
-
a Walker A box substitution in PilT, leads to abolishment of ATPase activity due to indirect interference withe ATP binding
-
K327A
-
a Walker A box substitution in PilB, leads to strong reduction of ATPase activity due to indirect interference withe ATP binding
-
D160N
-
mutation in the Walker A motif of PilT, and in PilU
D355N
-
mutation of PilB
E159Q
-
mutation in the Walker A motif of PilT, and in PilU
E163Q
-
mutation in the Walker A motif of PilT, and in PilU
E204
-
mutation in the Walker B motif of PilT, and in PilU, inactive PilT mutant
E354Q
-
mutation of PilB
E358Q
-
mutation of PilB
E396Q
-
mutation of PilB
G135S
-
mutation in the Asp Box of PilT, and in PilU, inactive PilT mutant
G331S
-
mutation of PilB
H222A
-
mutation in the His Box of PilT, and in PilU
H229A
-
mutation in the His Box of PilT, and in PilU
H414A
-
mutation of PilB
H421A
-
mutation of PilB
E383A
site-directed mutagenesis, the mutant shows almost unaltered protein secretion activity compared to the wild-type
E384A
site-directed mutagenesis, the mutant shows reduced protein secretion activity compared to the wild-type
E384A/Y385A
site-directed mutagenesis, the mutant shows reduced protein secretion activity compared to the wild-type
E384A/Y385A/G388A
site-directed mutagenesis, the mutant shows markedly reduced protein secretion activity and reduced ATPase activity compared to the wild-type
G164C
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loss-of-function mutant, unable to hydrolyze ATP, decrease in ability to interact with themselves and wild-type enzyme molecules
G388A
site-directed mutagenesis, the mutant shows slightly reduced protein secretion activity compared to the wild-type
K165E
site-directed mutagenesis, InvC ATPase inactive enzyme mutant
L376P
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loss-of-function mutant, defective in type II secretion and infection of cultured cells, wild-type ATP-ase activity
R189G
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loss-of-function mutant, unable to hydrolyze ATP
R191H
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loss-of-function mutant, unable to hydrolyze ATP, decrease in ability to interact with themselves and wild-type enzyme molecules
R192G
site-directed mutagenesis of the conserved arginine residue at position 192 of SsaN located in the dicyclohexylcarbodiimide-binding site in the catalytic domain, inactive mutant showing no ATPase and translocation activity. Introducing a plasmid that expresses SsaNR192G into the DELTAssaN mutant fails to complement SseJ secretion. Mutant SsaNR192G-Myc-His6 binds to chaperone SsaE
R223H
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loss-of-function mutant, unable to hydrolyze ATP
V28M
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loss-of-function mutant, defective in type II secretion and infection of cultured cells, wild-type ATP-ase activity
V51E
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loss-of-function mutant, defective in type II secretion and infection of cultured cells, wild-type ATP-ase activity
Y385A
site-directed mutagenesis, the Salmonelly typhimurium mutant strain expressing InvCY385A shows a marked defect in its ability to secrete the effector proteins SptP and SopB, while expression of the substrate proteins is unaffected by introduction of mutations in InvC, the mutant shows reduced ATPase activity compared to the wild-type
Y385W
site-directed mutagenesis, the mutant shows reduced ATPase activity compared to the wild-type
E383A
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site-directed mutagenesis, the mutant shows almost unaltered protein secretion activity compared to the wild-type
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E384A
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site-directed mutagenesis, the mutant shows reduced protein secretion activity compared to the wild-type
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K165E
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site-directed mutagenesis, InvC ATPase inactive enzyme mutant
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R192G
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site-directed mutagenesis of the conserved arginine residue at position 192 of SsaN located in the dicyclohexylcarbodiimide-binding site in the catalytic domain, inactive mutant showing no ATPase and translocation activity. Introducing a plasmid that expresses SsaNR192G into the DELTAssaN mutant fails to complement SseJ secretion. Mutant SsaNR192G-Myc-His6 binds to chaperone SsaE
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Y385A
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site-directed mutagenesis, the Salmonelly typhimurium mutant strain expressing InvCY385A shows a marked defect in its ability to secrete the effector proteins SptP and SopB, while expression of the substrate proteins is unaffected by introduction of mutations in InvC, the mutant shows reduced ATPase activity compared to the wild-type
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Y385W
-
site-directed mutagenesis, the mutant shows reduced ATPase activity compared to the wild-type
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K270A
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EpsE mutant
G175C
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catalytically inactive mutant
R286A
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XpsE mutant hydrolysis ATP at a rate five times that of the wild-type XpsE, but is non-functional in protein secretion via T2SS
K175E
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mutation in one of hte “Walker” boxes, enzymatically inactive
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged truncation mutant enzymes from Escherichia coli strain BL21(DE3) are unfolded under denaturing conditions in 6 M guanidium-HCl, 25 mM Tris pH 8.0, 1 mM EDTA, 300 mM NaCl, followed by dialysis and refolding in 25 mM Tris, 50 mM NaCl
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
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glutathione S-transferase-enzyme fusion protein functions analogously to native protein. Overexpression of regulatory protein YscL impairs secretion
drug development
medicine
pilD-dependent mechanism for promoting Legionella pneumophila intracellular infection of human cells
pharmacology