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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
|
| pubmed:dateCreated |
1996-11-4
|
| pubmed:abstractText |
An engineered interleukin-4-binding protein (IL4-BP) representing the extracellular domain of the human interleukin-4 (IL-4) receptor alpha chain was expressed in Sf9 cells. The purified IL4-BP was immobilized via a single biotinylated SH group near the carboxyl end to a biosensor matrix and analysed in real time for interaction with IL-4 and IL-4 variants. IL-4 was bound to IL4-BP at a molar ratio of approximately 1:1. The association and dissociation at pH 7.4 and 150 mM NaCl had rate constants of 1.9 +/- 0.3 x 10(7) M-1 s-1 and 2 +/- 1 x 10(-3) s-1, respectively. Glycosylation and engineered amino acid substitutions of IL4-BP did not alter the kinetic constants as shown by a parallel analysis of IL4-BP variants produced in Escherichia coli or Chinese hamster ovary cells. The rate of association was only slightly affected in binding-deficient variants [E9Q]IL-4 and [R88Q]IL-4 and by acidic pH down to values of 4.5, but it was reduced up to fivefold at higher ionic strength. The rate of dissociation was increased 70-fold and 150-fold with the IL-4 variants and fivefold at an acidic pH of 4.5, but it was not affected by high ionic strength. Temperatures between 6 degrees C and 37 degrees C yielded similar rates of IL-4 dissociation and only a marginally reduced rate of IL-4 association at 6 degrees C. These results indicate that the high-affinity binding of IL-4 to its receptor (Kd approximately 100 pM) is mainly the result of an unusually high association rate. The IL-4/IL4-BP interaction appears to be dominated by charge effects. The exceedingly high rate of IL-4/IL4-BP association is augmented by the overall electrostatic potentials of both proteins (electrostatic steering). Localized charges and the formation of ion pairs may control the rate of complex dissociation.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, CD,
http://linkedlifedata.com/resource/pubmed/chemical/Interleukin-4,
http://linkedlifedata.com/resource/pubmed/chemical/Macromolecular Substances,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Interleukin,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Interleukin-4,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Aug
|
| pubmed:issn |
0014-2956
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
15
|
| pubmed:volume |
240
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
252-61
|
| pubmed:dateRevised |
2008-11-21
|
| pubmed:meshHeading |
pubmed-meshheading:8797861-Animals,
pubmed-meshheading:8797861-Antigens, CD,
pubmed-meshheading:8797861-Biosensing Techniques,
pubmed-meshheading:8797861-CHO Cells,
pubmed-meshheading:8797861-Cell Line,
pubmed-meshheading:8797861-Cloning, Molecular,
pubmed-meshheading:8797861-Cricetinae,
pubmed-meshheading:8797861-Escherichia coli,
pubmed-meshheading:8797861-Genetic Variation,
pubmed-meshheading:8797861-Humans,
pubmed-meshheading:8797861-Hydrogen-Ion Concentration,
pubmed-meshheading:8797861-Interleukin-4,
pubmed-meshheading:8797861-Kinetics,
pubmed-meshheading:8797861-Macromolecular Substances,
pubmed-meshheading:8797861-Molecular Weight,
pubmed-meshheading:8797861-Mutagenesis, Site-Directed,
pubmed-meshheading:8797861-Osmolar Concentration,
pubmed-meshheading:8797861-Point Mutation,
pubmed-meshheading:8797861-Receptors, Interleukin,
pubmed-meshheading:8797861-Receptors, Interleukin-4,
pubmed-meshheading:8797861-Recombinant Proteins,
pubmed-meshheading:8797861-Spodoptera,
pubmed-meshheading:8797861-Thermodynamics,
pubmed-meshheading:8797861-Transfection
|
| pubmed:year |
1996
|
| pubmed:articleTitle |
Global and local determinants for the kinetics of interleukin-4/interleukin-4 receptor alpha chain interaction. A biosensor study employing recombinant interleukin-4-binding protein.
|
| pubmed:affiliation |
Theodor-Boveri-Institut für Biowissenschaften (Biozentrum) Universität Physiologische Chemie II, Würzburg, Germany.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|