Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
14
|
| pubmed:dateCreated |
1996-6-21
|
| pubmed:abstractText |
To determine the regions of interleukin-8 (IL-8) that allow high affinity and interleukin-8 receptor type 1 (IL8R1)-specific binding of chemokines, we produced chimeric proteins containing structural domains from IL-8, which binds to both IL8R1 and interleukin-8 receptor type 2 (IL8R2) with high affinity, and from GRO gamma, which does not bind to IL8R1 and binds to IL8R2 with reduced affinity. Receptor binding activity was tested by competition of 125I-IL-8 binding to recombinant IL8R1 and IL8R2 cell lines. Substitution into IL-8 of the GRO gamma sequences corresponding to either the amino-terminal loop (amino acids 1-18) or the first beta-sheet (amino acids 18-32) reduced binding to both IL8R1 and IL8R2. The third beta-sheet of IL-8 (amino acids 46-53) was required for binding to IL8R1 but not IL8R2. Exchanges of the second beta-sheet (amino acids 32-46) or the carboxyl-terminal alpha-helix (amino acids 53-72) had no significant effect. When IL-8 sequences were substituted into GRO gamma, a single domain containing the second beta-sheet of IL-8 (amino acids 18-32) was sufficient to confer high affinity binding for both IL8R1 and IL8R2. The amino-terminal loop (amino acids 1-18) and the third beta-sheet (amino acids 46-53) of IL-8 had little effect when substituted individually but showed increased binding to both receptors when substituted in combination. Individual amino acid substitutions were made at positions where IL-8 and GRO gamma sequences differ within the regions of residues 11-21 and 46-53. IL-8 mutations L49A or L49F selectively inhibited binding to IL8R1. Mutations Y13L and F21N enhanced binding to IL8R1 with little effect on IL8R2. A combined mutation Y13L/S14Q selectively decreased binding to IL8R2. Residues Tyr13, Ser14, Phe21, and Lys49 are clustered in and around a surface-accessible hydrophobic pocket on IL-8 that is physically distant from the previously identified ELR binding sequence. A homology model of GRO gamma, constructed from the known structure of IL-8 by refinement calculations, indicated that access to the hydrophobic pocket was effectively abolished in GRO gamma. These studies suggest that the surface hydrophobic pocket and/or adjacent residues participate in IL-8 receptor recognition for both IL8R1 and IL8R2 and that the hydrophobic pocket itself may be essential for IL8R1 binding. Thus this region contains a second site for IL-8 receptor recognition that, in combination with the Glu4-Leu5-Arg6 region, can modulate receptor binding affinity and IL8R1 specificity.
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, CD,
http://linkedlifedata.com/resource/pubmed/chemical/CXCL1 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Chemokine CXCL1,
http://linkedlifedata.com/resource/pubmed/chemical/Chemokines,
http://linkedlifedata.com/resource/pubmed/chemical/Chemokines, CXC,
http://linkedlifedata.com/resource/pubmed/chemical/Chemotactic Factors,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/Growth Substances,
http://linkedlifedata.com/resource/pubmed/chemical/Intercellular Signaling Peptides...,
http://linkedlifedata.com/resource/pubmed/chemical/Interleukin-8,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Interleukin,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Interleukin-8A,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Apr
|
| pubmed:issn |
0021-9258
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
5
|
| pubmed:volume |
271
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
8228-35
|
| pubmed:dateRevised |
2007-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:8626516-Amino Acid Sequence,
pubmed-meshheading:8626516-Antigens, CD,
pubmed-meshheading:8626516-Base Sequence,
pubmed-meshheading:8626516-Binding, Competitive,
pubmed-meshheading:8626516-Binding Sites,
pubmed-meshheading:8626516-Chemokine CXCL1,
pubmed-meshheading:8626516-Chemokines,
pubmed-meshheading:8626516-Chemokines, CXC,
pubmed-meshheading:8626516-Chemotactic Factors,
pubmed-meshheading:8626516-Chemotaxis, Leukocyte,
pubmed-meshheading:8626516-DNA Primers,
pubmed-meshheading:8626516-Growth Substances,
pubmed-meshheading:8626516-Humans,
pubmed-meshheading:8626516-Intercellular Signaling Peptides and Proteins,
pubmed-meshheading:8626516-Interleukin-8,
pubmed-meshheading:8626516-Models, Molecular,
pubmed-meshheading:8626516-Molecular Sequence Data,
pubmed-meshheading:8626516-Neutrophils,
pubmed-meshheading:8626516-Protein Binding,
pubmed-meshheading:8626516-Protein Structure, Tertiary,
pubmed-meshheading:8626516-Receptors, Interleukin,
pubmed-meshheading:8626516-Receptors, Interleukin-8A,
pubmed-meshheading:8626516-Recombinant Fusion Proteins,
pubmed-meshheading:8626516-Recombinant Proteins,
pubmed-meshheading:8626516-Sequence Alignment,
pubmed-meshheading:8626516-Sequence Homology, Amino Acid,
pubmed-meshheading:8626516-Solubility
|
| pubmed:year |
1996
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| pubmed:articleTitle |
Receptor recognition and specificity of interleukin-8 is determined by residues that cluster near a surface-accessible hydrophobic pocket.
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| pubmed:affiliation |
Chiron Corporation, Emeryville, California 94608, USA.
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| pubmed:publicationType |
Journal Article
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