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Information on EC 7.6.2.9 - ABC-type quaternary amine transporter and Organism(s) Bacillus subtilis and UniProt Accession P46920

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IUBMB Comments
An ATP-binding cassette (ABC) type transporter, characterized by the presence of two similar ATP-binding domains/proteins and two integral membrane domains/proteins. Does not undergo phosphorylation during the transport process. A bacterial enzyme that interacts with an extracytoplasmic substrate binding protein and mediates the high affinity uptake of quaternary amine derivatives.
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Bacillus subtilis
UNIPROT: P46920
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Word Map
The taxonomic range for the selected organisms is: Bacillus subtilis
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
+
+
quaternary amine-[quaternary amine-binding protein][side 1]
=
+
+
quaternary amine[side 2]
+
[quaternary amine-binding protein][side 1]
Synonyms
abc transporter, opuac, glycine betaine-binding protein, opubc, opucc, glycine betaine porter ii, opuab, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ABC transporter
-
glycine betaine transport system permease protein
glycine betaine-binding protein
-
glycine betaine-binding protein OpuAC
UniProt
glycine betaine/carnitine/choline-binding protein
-
Quaternary-amine-transporting ATPase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + H2O + quaternary amine-[quaternary amine-binding protein][side 1] = ADP + phosphate + quaternary amine[side 2] + [quaternary amine-binding protein][side 1]
show the reaction diagram
OpuA is an osmoprotectant uptake system which imports glycine and betaine. It consists of three components, the ATPase OpuAA, the integral membrane protein OpuAB and the extracellular substrate binding domain OpuAC. The ABC transporter couples ATP hydrolysis with substrate translocation across the membrane in a vectorial manner
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase (ABC-type, quaternary-amine-importing)
An ATP-binding cassette (ABC) type transporter, characterized by the presence of two similar ATP-binding domains/proteins and two integral membrane domains/proteins. Does not undergo phosphorylation during the transport process. A bacterial enzyme that interacts with an extracytoplasmic substrate binding protein and mediates the high affinity uptake of quaternary amine derivatives.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-83-3
-
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
ADP + H2O
AMP + phosphate
show the reaction diagram
-
OpuAA protein, ATPase activity
-
-
?
AMP + H2O
adenine + phosphate
show the reaction diagram
-
OpuAA protein, ATPase activity, similar substrate affinities to monomeric and dimeric states
-
-
?
ATP + H2O
ADP + phosphate
show the reaction diagram
ATP + H2O + betaine/out
ADP + phosphate + betaine/in
show the reaction diagram
-
-
-
-
?
ATP + H2O + betonicine/out
ADP + phosphate + betonicine/in
show the reaction diagram
-
-
-
-
?
ATP + H2O + dimethylsulfonioacetate/out
ADP + phosphate + dimethylsulfonioacetate/in
show the reaction diagram
ATP + H2O + glycine betaine/out
ADP + phosphate + glycine betaine/in
show the reaction diagram
ATP + H2O + L-proline betaine/out
ADP + phosphate + L-proline betaine/in
show the reaction diagram
-
-
-
-
?
ATP + H2O + proline betaine/out
ADP + phosphate + proline betaine/in
show the reaction diagram
-
-
-
-
?
ATP + H2O + quaternary amine-[quaternary amine-binding protein][side 1]
ADP + phosphate + quaternary amine[side 2] + [quaternary amine-binding protein][side 1]
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL 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
-
-
-
?
ATP + H2O + dimethylsulfonioacetate/out
ADP + phosphate + dimethylsulfonioacetate/in
show the reaction diagram
OpuA is a classic ABC importer that relies on a substrate binding protein priming the transporter with specificity and selectivity
-
-
?
ATP + H2O + glycine betaine/out
ADP + phosphate + glycine betaine/in
show the reaction diagram
ATP + H2O + quaternary amine-[quaternary amine-binding protein][side 1]
ADP + phosphate + quaternary amine[side 2] + [quaternary amine-binding protein][side 1]
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
-
K+ binding site within the nucleotide-binding protein domain, stimulation of the nucleotide-binding protein domain 4fold higher than with Na+
Na+
-
stimulation of the nucleotide-binding protein domain 4fold lower than with K+
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
nucleotide-binding protein domain not stimulated by glycine betaine
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.6 - 2.8
ATP
0.25 - 5.4
ATP
0.0024 - 0.013
glycine betaine
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.004 - 0.25
ATP
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 24
assay at
37 - 52
-
reduced growth rate at higher temperatures, addition of 1 mM glycine betaine, homobetaine, carnitine, butyrobetaine, crotonobetaine, DMSP, choline, or glutamate have a clear thermoprotective effect at 52°C, heat protectants are taken up under heat stress via the OpuA, OpuC, and OpuD transporters, despite the strongly reduced glycine betaine transport rate at 52°C, substantial glycine betaine accumulation, but it is not increased in comparison to cells grown at 37°C, thermoprotection by glutamate does not depend on an increased cellular pool of this amino acid
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
SwissProt
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
substrate-binding protein domain
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28000
-
substrate-binding domain, 1 * 28000, SDS-PAGE
47700
-
OpuAA monomer, calculated from cDNA
56130
-
x * 56130, calculation from nucleotide sequence
65000
-
OpuAA, gel filtration
95400
-
OpuAA dimer, calculated from cDNA
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 56130, calculation from nucleotide sequence
dimer
-
nucleotide-binding protein domain, displays lower ATPase activity than the dimeric form
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
lipoprotein
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
by hanging-drop method, structures of substrate-binding domain in complex with glycine betaine solved at 2.0 A resolution and in complex with glycine proline at 2.8 A resolution, structures show a substrate-binding protein-dependent-typical class II fold, structural differences of complexes occur within the ligand-binding pocket as well as across the domain-domain interface, explaining the differences in affinity of the substrate-binding domain-glycine betaine complex with KD = 0.017 mM, and substrate-binding domain-proline betaine complex with KD = 0.295 mM
-
OpuBC protein crystal structure analysis, PDB ID 3R6U
substrate-binding domain in complex with glycine betaine solved at 2.0 A resolution and in complex with glycine proline at 2.8 A resolution
-
substrate-binding protein OpuBC in complex with choline, to 1.6 A resolution. The positively charged trimethylammonium head group of choline is wedged into an aromatic cage formed by four tyrosine residues and is bound via cation-pi interactions. The hydroxyl group of choline protrudes out of this aromatic cage and makes a single interaction with residue Gln19. A water network stabilizes choline within its substrate-binding site and promotes indirect interactions between the two lobes of the OpuBC protein. Disruption of this intricate water network strongly reduces choline binding affinity or abrogates ligand binding
substrate-binding protein OpuCC in the apo-form and in complex with carnitine, glycine betaine, choline and ectoine to 2.3, 2.7, 2.4, 1.9 and 2.1 A resolution, respectively. OpuCC is composed of two alpha/beta/alpha globular sandwich domains linked by two hinge regions, with a substrate-binding pocket located at the interdomain cleft. Upon substrate binding, the two domains shift towards each other to trap the substrate
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F19W
mutant analyzed
G161C
single cysteine mutants generated by site-directed mutagenesis
S171C
single cysteine mutants generated by site-directed mutagenesis
S45C
single cysteine mutants generated by site-directed mutagenesis
D149A
decrease in choline binding affinity by approximately 18fold
D149A/L155A
decrease in choline binding affinity by approximately 38fold
D74A
mutant is unable to bind choline
E171Q
-
the monomer is the preferred species for the nucleotide-free state in solution
L155A
decrease in choline binding affinity by approximately 25fold
M21A
decrease in choline binding affinity by approximately 3fold
N115A
mutant is unable to bind choline
Q19A
decrease in choline binding affinity by approximately 15fold
Q19A/L155A
mutant is unable to bind choline
T94D
shares a quite similar pattern of fluorescence spectrum to that of the paralogue OpuBC. Only choline can trigger obvious changes of fluorescence intensity of mutant T94D, whereas carnitine, GB and ectoine cannot
Y91A
complete loss of binding affinity
Y91F
slight decrease in binding affinity
Y91W
slight decrease in binding affinity
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
both monomeric and dimeric OpuA species are stabilized at 1 M NaCl
-
OpuAA forms a stable dimer in the nucleotide-free state in solution
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
by affinity tag as first step and gel filtration
-
isolated transmembrane subunits purified by Strep-tag affinity purification
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OpuAA, purified to homogeneity, affinity chromatography and gel filtration
-
substrate-binding domain, to apparent homogeneity
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli, cysteine mutants generated and inserted into pBAD33
expressed in Escherichia coli BL21 cells
-
expression in Escherichia coli
expression of OpuAA protein in Escherichia coli
-
gene opuAB, sequence comparisons
gene opuAC, sequence comparisons
nucleotide-binding protein domain cloned as an N- or C-terminal His-tagged fusion protein and overexpressed in Escherichia coli BL21 (DE3) under the control of an arabinose-inducible promoter, overexpression of the transmembrane domain only possible in Walker strains BL21(DE3)C41 and C43 under the control of an ITPG- or arabinose-inducible promoter as N- and C-terminal His- and Strep-tagged versions
-
substrate-binding domain into vector pBKB76, expression in Escherichia coli BL21 (DE3)
-
transmembrane subunits cloned into vector pBAD33 or pET21a, expression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enzyme expression is induced in response to an osmotic upshock and sustained high salinity
-
the MarR-type regulator GbsR, which acts as an intracellular choline-responsive repressor, controls the transcription of the opuB and gbsAB operons
the trans-acting regulatory gene gbsR encodes a repressor (GbsR) for the opuB operon
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Van der Heide, T.; Poolman, B.
Osmoregulated ABC-transport system of Lactococcus lactis senses water stress via changes in the physical state of the membrane
Proc. Natl. Acad. Sci. USA
97
7102-7106
2000
Bacillus subtilis, Lactococcus lactis, Lactococcus lactis OpuA
Manually annotated by BRENDA team
Kempf, B.; Bremer, E.
OpuA, an osmotically regulated binding protein-dependent transport system for the osmoprotectant glycine betaine in Bacillus subtilis
J. Biol. Chem.
270
16701-16713
1995
Bacillus subtilis, Bacillus subtilis OpuA
Manually annotated by BRENDA team
Kempf, B.; Gade, J.; Bremer, E.
Lipoprotein from the osmoregulated ABC transport system OpuA of Bacillus subtilis: purification of the glycine betaine binding protein and characterization of a functional lipidless mutant
J. Bacteriol.
179
6213-6220
1997
Bacillus subtilis
Manually annotated by BRENDA team
Kappes, R.M.; Kempf, B.; Bremer, E.
Three transport systems for the osmoprotectant glycine betaine operate in Bacillus subtilis: characterization of OpuD
J. Bacteriol.
178
5071-5079
1996
Bacillus subtilis
Manually annotated by BRENDA team
Horn, C.; Bremer, E.; Schmitt, L.
Nucleotide dependent monomer/dimer equilibrium of OpuAA, the nucleotide-binding protein of the osmotically regulated ABC transporter OpuA from Bacillus subtilis
J. Mol. Biol.
334
403-419
2003
Bacillus subtilis
Manually annotated by BRENDA team
Horn, C.; Bremer, E.; Schmitt, L.
Functional overexpression and in vitro re-association of OpuA, an osmotically regulated ABC-transport complex from Bacillus subtilis
FEBS Lett.
579
5765-5768
2005
Bacillus subtilis
Manually annotated by BRENDA team
Holtmann, G.; Bremer, E.
Thermoprotection of Bacillus subtilis by exogenously provided glycine betaine and structurally related compatible solutes: involvement of Opu transporters
J. Bacteriol.
186
1683-1693
2004
Bacillus subtilis
Manually annotated by BRENDA team
Horn, C.; Sohn-Boesser, L.; Breed, J.; Welte, W.; Schmitt, L.; Bremer, E.
Molecular determinants for substrate specificity of the ligand-binding protein OpuAC from Bacillus subtilis for the compatible solutes glycine betaine and proline betaine
J. Mol. Biol.
357
592-606
2006
Bacillus subtilis
Manually annotated by BRENDA team
Horn, C.; Jenewein, S.; Sohn-Boesser, L.; Bremer, E.; Schmitt, L.
Biochemical and structural analysis of the Bacillus subtilis ABC transporter OpuA and its isolated subunits
J. Mol. Microbiol. Biotechnol.
10
76-91
2005
Bacillus subtilis, Lactococcus lactis
Manually annotated by BRENDA team
Horn, C.; Jenewein, S.; Tschapek, B.; Bouschen, W.; Metzger, S.; Bremer, E.; Schmitt, L.
Monitoring conformational changes during the catalytic cycle of OpuAA, the ATPase subunit of the ABC transporter OpuA from Bacillus subtilis
Biochem. J.
412
233-244
2008
Bacillus subtilis (P46920), Bacillus subtilis
Manually annotated by BRENDA team
Smits, S.H.; Hoeing, M.; Lecher, J.; Jebbar, M.; Schmitt, L.; Bremer, E.
The compatible-solute-binding protein OpuAC from Bacillus subtilis: ligand binding, site-directed mutagenesis, and crystallographic studies
J. Bacteriol.
190
5663-5671
2008
Bacillus subtilis (P46922), Bacillus subtilis
Manually annotated by BRENDA team
Du, Y.; Shi, W.W.; He, Y.X.; Yang, Y.H.; Zhou, C.Z.; Chen, Y.
Structures of the substrate-binding protein provide insights into the multiple compatible solute binding specificities of the Bacillus subtilis ABC transporter OpuC
Biochem. J.
436
283-289
2011
Bacillus subtilis (O32243), Bacillus subtilis
Manually annotated by BRENDA team
Pittelkow, M.; Tschapek, B.; Smits, S.H.; Schmitt, L.; Bremer, E.
The crystal structure of the substrate-binding protein OpuBC from Bacillus subtilis in complex with choline
J. Mol. Biol.
411
53-67
2011
Bacillus subtilis (Q45462), Bacillus subtilis, Bacillus subtilis JH642 (Q45462)
Manually annotated by BRENDA team
Hoffmann, T.; Wensing, A.; Brosius, M.; Steil, L.; Voelker, U.; Bremer, E.
Osmotic control of opuA expression in Bacillus subtilis and its modulation in response to intracellular glycine betaine and proline pools
J. Bacteriol.
195
510-522
2013
Bacillus subtilis, Bacillus subtilis JH642
Manually annotated by BRENDA team
Bashir, A.; Hoffmann, T.; Kempf, B.; Xie, X.; Smits, S.; Bremer, E.
Plant-derived compatible solutes proline betaine and betonicine confer enhanced osmotic and temperature stress tolerance to Bacillus subtilis
Microbiology
160
2283-2294
2014
Bacillus subtilis
Manually annotated by BRENDA team
Ronzheimer, S.; Warmbold, B.; Arnhold, C.; Bremer, E.
The GbsR family of transcriptional regulators functional characterization of the OpuAR repressor
Front. Microbiol.
9
2536
2018
Bacillus subtilis (P46921), Bacillus subtilis (P46922), Bacillus infantis (U5LFB4 AND U5LHK3 AND U5LHN4), Bacillus subtilis 168 (P46921), Bacillus subtilis 168 (P46922), Bacillus infantis NRRL B-14911 (U5LFB4 AND U5LHK3 AND U5LHN4)
Manually annotated by BRENDA team
Teichmann, L.; Chen, C.; Hoffmann, T.; Smits, S.H.J.; Schmitt, L.; Bremer, E.
From substrate specificity to promiscuity hybrid ABC transporters for osmoprotectants
Mol. Microbiol.
104
761-780
2017
Bacillus subtilis (P46921), Bacillus subtilis (P46922), Bacillus subtilis, Bacillus subtilis 168 (P46921), Bacillus subtilis 168 (P46922), Bacillus subtilis JH642 (P46921), Bacillus subtilis JH642 (P46922)
Manually annotated by BRENDA team