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PDBsum entry 1y3q
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Sugar binding protein
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PDB id
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1y3q
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References listed in PDB file
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Key reference
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Title
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Direct evidence for sphingomonas sp. A1 periplasmic proteins as macromolecule-Binding proteins associated with the abc transporter: molecular insights into alginate transport in the periplasm.
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Authors
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K.Momma,
Y.Mishima,
W.Hashimoto,
B.Mikami,
K.Murata.
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Ref.
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Biochemistry, 2005,
44,
5053-5064.
[DOI no: ]
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PubMed id
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Abstract
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A Gram-negative bacterium, Sphingomonas sp. A1, has a macromolecule (alginate)
import system consisting of a pit on the cell surface and an alginate-specific
ATP-binding cassette importer in the inner membrane. Transport of alginate from
the pit to the ABC importer is probably mediated by two periplasmic binding
protein homologues (AlgQ1 and AlgQ2). Here we describe characteristics of
binding of AlgQ1 and AlgQ2 to alginate and its oligosaccharides through surface
plasmon resonance biosensor analysis, UV absorption difference spectroscopy, and
X-ray crystallography. Both AlgQ1 and AlgQ2 were inducibly expressed in the
periplasm of alginate-grown cells of strain A1. Biosensor analysis indicated
that both proteins specifically bind alginate with a high degree of
polymerization (>100) and that dissociation constants for alginate with an
average molecular mass of 26 kDa are 2.3 x 10(-)(7) M for AlgQ1 and 1.5 x
10(-)(7) M for AlgQ2. An in vitro ATPase assay using the membrane complex,
including the alginate ABC importer, suggested that both alginate-bound forms of
AlgQ1 and AlgQ2 are closely associated with the importer. X-ray crystallography
showed that AlgQ1 consisted of two domains separated by a deep cleft that binds
alginate oligosaccharides through a conformational change in the two domains.
These results directly show that alginate-binding proteins play an important
role in the efficient transport of alginate macromolecules with different
degrees of polymerization in the periplasm.
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