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PDBsum entry 3d9b
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Transport protein
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PDB id
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3d9b
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References listed in PDB file
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Key reference
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Title
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There is a baby in the bath water: acrb contamination is a major problem in membrane-Protein crystallization.
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Authors
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D.Veesler,
S.Blangy,
C.Cambillau,
G.Sciara.
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Ref.
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Acta Crystallogr Sect F Struct Biol Cryst Commun, 2008,
64,
880-885.
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PubMed id
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Abstract
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In the course of a crystallographic study of the Methanosarcina mazei CorA
transporter, the membrane protein was obtained with at least 95% purity and was
submitted to crystallization trials. Small crystals (<100 microm) were grown
that diffracted to 3.42 A resolution and belonged to space group R32, with
unit-cell parameters a = b = 145.74, c = 514.0 A. After molecular-replacement
attempts using available CorA structures as search models failed to yield a
solution, it was discovered that the crystals consisted of an Escherichia coli
contaminating protein, acriflavine resistance protein B (AcrB), that was present
at less than 5% in the protein preparations. AcrB contamination is a major
problem when expressing membrane proteins in E. coli since it binds naturally to
immobilized metal-ion affinity chromatography (IMAC) resins. Here, the structure
is compared with previously deposited AcrB structures and strategies are
proposed to avoid this contamination.
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