Source:http://linkedlifedata.com/resource/pubmed/id/16489764
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
8
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| pubmed:dateCreated |
2006-2-21
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| pubmed:abstractText |
The cell surface receptor for bacteriophage Lambda is LamB (maltoporin). Responsible for phage binding to LamB is the C-terminal part, gpJ, of phage tail protein J. To study the interaction between LamB and gpJ, a chimera protein composed of maltose binding protein (MBP or MalE) connected to the C-terminal part of J (gpJ, amino acids 684-1131) of phage tail protein J of bacteriophage Lambda was expressed in Escherichia coli and purified to homogeneity. The interaction of the MBP-gpJ chimera protein with reconstituted LamB and its mutants LamB Y118G and the loop deletion mutant LamB Delta4+Delta6+Delta9v was studied using planar lipid bilayer membranes on a single-channel and multichannel level. Titration with the MBP-gpJ chimera blocked completely the ion current through reconstituted LamB when it was added to the cis side, the extracellular side of LamB with a half-saturation constant of approximately 6 nM in 1 M KCl. Control experiments with LamB Delta4+Delta6+Delta9v from which all major external loops had been removed showed similar blocking, whereas MBP alone caused no visible effect. Direct conductance measurement with His(6)-gpJ that contained a hexahistidyl tag (His(6) tag) at the N-terminal end of the protein for easy purification revealed no blocking of the ion current, requiring other measurements for the binding constant. However, when maltoporin was preincubated with His-gpJ, MBP-gpJ could not block the channel, which indicated that also His(6)-gpJ bound to the channel. High-molecular mass bands on SDS-PAGE and Western blots, confirming the planar lipid bilayer experiment results, also demonstrated stable complex formation between His(6)-gpJ and LamB or LamB mutants. The results revealed that phage Lambda binding includes not only the extracellular loops.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Outer Membrane Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Edetic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Histidine,
http://linkedlifedata.com/resource/pubmed/chemical/J protein, Bacteriophage lambda,
http://linkedlifedata.com/resource/pubmed/chemical/Magnesium,
http://linkedlifedata.com/resource/pubmed/chemical/Maltose-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Oligosaccharides,
http://linkedlifedata.com/resource/pubmed/chemical/Porins,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Cell Surface,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Virus,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Viral Tail Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/maltopentaose,
http://linkedlifedata.com/resource/pubmed/chemical/maltoporins
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| pubmed:status |
MEDLINE
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| pubmed:month |
Feb
|
| pubmed:issn |
0006-2960
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
28
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| pubmed:volume |
45
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
2708-20
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| pubmed:dateRevised |
2010-11-18
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| pubmed:meshHeading |
pubmed-meshheading:16489764-Bacterial Outer Membrane Proteins,
pubmed-meshheading:16489764-Bacteriophage lambda,
pubmed-meshheading:16489764-Blotting, Western,
pubmed-meshheading:16489764-Carrier Proteins,
pubmed-meshheading:16489764-Edetic Acid,
pubmed-meshheading:16489764-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:16489764-Enzyme Stability,
pubmed-meshheading:16489764-Gene Deletion,
pubmed-meshheading:16489764-Histidine,
pubmed-meshheading:16489764-Ion Transport,
pubmed-meshheading:16489764-Magnesium,
pubmed-meshheading:16489764-Maltose-Binding Proteins,
pubmed-meshheading:16489764-Mutation,
pubmed-meshheading:16489764-Oligosaccharides,
pubmed-meshheading:16489764-Porins,
pubmed-meshheading:16489764-Protein Binding,
pubmed-meshheading:16489764-Receptors, Cell Surface,
pubmed-meshheading:16489764-Receptors, Virus,
pubmed-meshheading:16489764-Recombinant Fusion Proteins,
pubmed-meshheading:16489764-Viral Tail Proteins
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| pubmed:year |
2006
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| pubmed:articleTitle |
Interaction of bacteriophage lambda with its cell surface receptor: an in vitro study of binding of the viral tail protein gpJ to LamB (Maltoporin).
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| pubmed:affiliation |
Lehrstuhl für Biotechnologie, Biocenter of the University of Würzburg, Am Hubland, D-97074 Würzburg, Federal Republic of Germany.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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