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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
3
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pubmed:dateCreated |
2001-5-8
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pubmed:abstractText |
Novel antagonists of the proinflammatory leukocyte chemoattractant C5a have been produced from a phage display library of whole-molecule random mutants. The cDNA for the inflammatory polypeptide C5adR(74) was used as template in a PCR reaction doped with the mutagenic nucleoside triphosphates dPTP [dP: 6-(2-deoxy-beta-D-ribofuranosyl)-3,4-dihydro-8H-pyrimido-[4,5-c][1,2]oxazin-7-one] and 8-oxodGTP (8-oxodG: 8-oxo-2'-deoxyguanosine) to allow the introduction of mutations in a highly controlled manner throughout the cDNA. The resultant library of mutants was displayed on bacteriophage M13 using a jun/fos linker sequence. Functional polypeptides were isolated by several rounds of selection against the receptor for C5a expressed on the surface of CHO cells. From this selection procedure, a limited number of variants of C5adR(74) were obtained. When expressed as free polypeptide, the binding affinities of the selected C5adR(74) sequences were increased 5-fold relative to wild-type protein. Site-directed mutagenesis of the C-terminus of these variants resulted in the production of antagonists of C5adR(74) activity.
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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 |
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, CD,
http://linkedlifedata.com/resource/pubmed/chemical/Complement C5a,
http://linkedlifedata.com/resource/pubmed/chemical/Ligands,
http://linkedlifedata.com/resource/pubmed/chemical/Nucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Receptor, Anaphylatoxin C5a,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Cell Surface,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Complement,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0269-2139
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
14
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
189-93
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:11342716-Amino Acid Sequence,
pubmed-meshheading:11342716-Amino Acid Substitution,
pubmed-meshheading:11342716-Animals,
pubmed-meshheading:11342716-Antigens, CD,
pubmed-meshheading:11342716-Bacteriophage M13,
pubmed-meshheading:11342716-Binding, Competitive,
pubmed-meshheading:11342716-Binding Sites,
pubmed-meshheading:11342716-CHO Cells,
pubmed-meshheading:11342716-Cell Line,
pubmed-meshheading:11342716-Complement C5a,
pubmed-meshheading:11342716-Cricetinae,
pubmed-meshheading:11342716-Gene Library,
pubmed-meshheading:11342716-Genetic Variation,
pubmed-meshheading:11342716-Humans,
pubmed-meshheading:11342716-Ligands,
pubmed-meshheading:11342716-Mutagenesis,
pubmed-meshheading:11342716-Mutagenesis, Site-Directed,
pubmed-meshheading:11342716-Nucleotides,
pubmed-meshheading:11342716-Polymerase Chain Reaction,
pubmed-meshheading:11342716-Proteins,
pubmed-meshheading:11342716-Receptor, Anaphylatoxin C5a,
pubmed-meshheading:11342716-Receptors, Cell Surface,
pubmed-meshheading:11342716-Receptors, Complement,
pubmed-meshheading:11342716-Recombinant Proteins,
pubmed-meshheading:11342716-Structure-Activity Relationship,
pubmed-meshheading:11342716-Transfection
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pubmed:year |
2001
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pubmed:articleTitle |
Selection of novel ligands from a whole-molecule randomly mutated C5a library.
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pubmed:affiliation |
Krebs Institute of Biomolecular Science, Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2UH, UK.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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