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PDBsum entry 3ejz
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Immune system/proton transport
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PDB id
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3ejz
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Contents |
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444 a.a.
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221 a.a.
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211 a.a.
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* Residue conservation analysis
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PDB id:
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| Name: |
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Immune system/proton transport
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Title:
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Structure of e203v mutant e.Coli cl-/h+ exchanger, clc-ec1
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Structure:
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H(+)/cl(-) exchange transporter clca. Chain: a, b. Synonym: clc-ec1 h(+)/cl(-) exchange. Clc-ec1. Engineered: yes. Mutation: yes. Fab fragment, heavy chain. Chain: c, e. Fab fragment, light chain. Chain: d, f
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Source:
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Escherichia coli. Organism_taxid: 562. Gene: clca, eric, yadq, b0155, jw5012. Expressed in: escherichia coli. Expression_system_taxid: 562. Mus musculus. Mouse. Organism_taxid: 10090. Cell: hybridoma.
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Resolution:
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2.90Å
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R-factor:
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0.253
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R-free:
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0.281
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Authors:
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H.-H.Lim,C.Miller
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Key ref:
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H.H.Lim
and
C.Miller
(2009).
Intracellular proton-transfer mutants in a CLC Cl-/H+ exchanger.
J Gen Physiol,
133,
131-138.
PubMed id:
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Date:
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18-Sep-08
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Release date:
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17-Feb-09
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PROCHECK
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Headers
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References
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P37019
(CLCA_ECOLI) -
H(+)/Cl(-) exchange transporter ClcA from Escherichia coli (strain K12)
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Seq:
Struc:
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473 a.a.
444 a.a.*
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J Gen Physiol
133:131-138
(2009)
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PubMed id:
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Intracellular proton-transfer mutants in a CLC Cl-/H+ exchanger.
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H.H.Lim,
C.Miller.
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ABSTRACT
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CLC-ec1, a bacterial homologue of the CLC family's transporter subclass,
catalyzes transmembrane exchange of Cl(-) and H(+). Mutational analysis based on
the known structure reveals several key residues required for coupling H(+) to
the stoichiometric countermovement of Cl(-). E148 (Glu(ex)) transfers protons
between extracellular water and the protein interior, and E203 (Glu(in)) is
thought to function analogously on the intracellular face of the protein.
Mutation of either residue eliminates H(+) transport while preserving Cl(-)
transport. We tested the role of Glu(in) by examining structural and functional
properties of mutants at this position. Certain dissociable side chains (E, D,
H, K, R, but not C and Y) retain H(+)/Cl(-) exchanger activity to varying
degrees, while other mutations (V, I, or C) abolish H(+) coupling and severely
inhibit Cl(-) flux. Transporters substituted with other nonprotonatable side
chains (Q, S, and A) show highly impaired H(+) transport with substantial Cl(-)
transport. Influence on H(+) transport of side chain length and acidity was
assessed using a single-cysteine mutant to introduce non-natural side chains.
Crystal structures of both coupled (E203H) and uncoupled (E203V) mutants are
similar to wild type. The results support the idea that Glu(in) is the internal
proton-transfer residue that delivers protons from intracellular solution to the
protein interior, where they couple to Cl(-) movements to bring about Cl(-)/H(+)
exchange.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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G.V.Miloshevsky,
A.Hassanein,
and
P.C.Jordan
(2010).
Antiport mechanism for Cl(-)/H(+) in ClC-ec1 from normal-mode analysis.
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Biophys J,
98,
999.
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L.Feng,
E.B.Campbell,
Y.Hsiung,
and
R.MacKinnon
(2010).
Structure of a eukaryotic CLC transporter defines an intermediate state in the transport cycle.
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Science,
330,
635-641.
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PDB code:
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L.Wellhauser,
C.D'Antonio,
and
C.E.Bear
(2010).
ClC transporters: discoveries and challenges in defining the mechanisms underlying function and regulation of ClC-5.
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Pflugers Arch,
460,
543-557.
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R.J.Naftalin
(2010).
Reassessment of models of facilitated transport and cotransport.
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J Membr Biol,
234,
75.
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A.Picollo,
M.Malvezzi,
J.C.Houtman,
and
A.Accardi
(2009).
Basis of substrate binding and conservation of selectivity in the CLC family of channels and transporters.
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Nat Struct Mol Biol,
16,
1294-1301.
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D.Wang,
and
G.A.Voth
(2009).
Proton transport pathway in the ClC Cl-/H+ antiporter.
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Biophys J,
97,
121-131.
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E.Y.Bergsdorf,
A.A.Zdebik,
and
T.J.Jentsch
(2009).
Residues important for nitrate/proton coupling in plant and mammalian CLC transporters.
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J Biol Chem,
284,
11184-11193.
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H.H.Lim,
and
C.Miller
(2009).
Intracellular proton-transfer mutants in a CLC Cl-/H+ exchanger.
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J Cell Biol,
184,
131-138.
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The most recent references are shown first.
Citation data come partly from CiteXplore and partly
from an automated harvesting procedure. Note that this is likely to be
only a partial list as not all journals are covered by
either method. However, we are continually building up the citation data
so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
code is
shown on the right.
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Links
