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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2009-3-2
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pubmed:databankReference |
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pubmed:abstractText |
Pseudopilins form the central pseudopilus of the sophisticated bacterial type 2 secretion systems. The crystallization of the EpsI:EpsJ pseudopilin heterodimer from Vibrio vulnificus was greatly accelerated by the use of nanobodies, which are the smallest antigen-binding fragments derived from heavy-chain only camelid antibodies. Seven anti-EpsI:EpsJ nanobodies were generated and co-crystallization of EpsI:EpsJ nanobody complexes yielded several crystal forms very rapidly. In the structure solved, the nanobodies are arranged in planes throughout the crystal lattice, linking layers of EpsI:EpsJ heterodimers. The EpsI:EpsJ dimer observed confirms a right-handed architecture of the pseudopilus, but, compared to a previous structure of the EpsI:EpsJ heterodimer, EpsI differs 6 degrees in orientation with respect to EpsJ; one loop of EpsJ is shifted by approximately 5A due to interactions with the nanobody; and a second loop of EpsJ underwent a major change of 17A without contacts with the nanobody. Clearly, nanobodies accelerate dramatically the crystallization of recalcitrant protein complexes and can reveal conformational flexibility not observed before.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
1095-8657
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:volume |
166
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
8-15
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pubmed:dateRevised |
2011-11-17
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pubmed:meshHeading |
pubmed-meshheading:19118632-Amino Acid Sequence,
pubmed-meshheading:19118632-Animals,
pubmed-meshheading:19118632-Antibodies, Monoclonal,
pubmed-meshheading:19118632-Antigen-Antibody Complex,
pubmed-meshheading:19118632-Binding Sites, Antibody,
pubmed-meshheading:19118632-Camelids, New World,
pubmed-meshheading:19118632-Complementarity Determining Regions,
pubmed-meshheading:19118632-Crystallization,
pubmed-meshheading:19118632-Crystallography, X-Ray,
pubmed-meshheading:19118632-Epitopes,
pubmed-meshheading:19118632-Immunoglobulin Heavy Chains,
pubmed-meshheading:19118632-Membrane Transport Proteins,
pubmed-meshheading:19118632-Models, Molecular,
pubmed-meshheading:19118632-Molecular Sequence Data,
pubmed-meshheading:19118632-Protein Multimerization,
pubmed-meshheading:19118632-Protein Structure, Quaternary,
pubmed-meshheading:19118632-Recombinant Proteins,
pubmed-meshheading:19118632-Sequence Alignment,
pubmed-meshheading:19118632-Vibrio vulnificus
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pubmed:year |
2009
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pubmed:articleTitle |
Nanobody-aided structure determination of the EpsI:EpsJ pseudopilin heterodimer from Vibrio vulnificus.
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pubmed:affiliation |
Department of Biochemistry, Biomolecular Structure Center, University of Washington, 1959 Pacific Ave. NE, HSC K-428, Seattle, WA 98195, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
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