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rdf:type |
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lifeskim:mentions |
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pubmed:dateCreated |
1999-6-30
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pubmed:abstractText |
The placement of a tryptophan residue into a single Ig-binding-domain of protein L from Peptostreptococcus magnus has been used to examine the binding interactions between the binding domain and kappa light chains (kappa-chains). The fluorescence intensity of the mutant domain increases on the formation of a complex with kappa-chains. This has been used to determine the Kd of the complex under a range of conditions by using both pre-equilibrium and equilibrium methods. The Kd values determined for the complex with kappa-chains at a number of different pH values are very close to those obtained with the wild-type domain, indicating that the mutation has not substantially affected its binding properties. Examination of the reaction between the mutant domain and kappa-chains by stopped-flow fluorescence shows that complex formation takes place by two discrete, sequential processes. A fast bimolecular reaction, with a rate constant of 8.3x10(5) M-1. s-1 (at pH8.0 and 25 degrees C), is followed by a slow unimolecular process with a rate (1.45 s-1) that is independent of the concentration of the reactants. This suggests that a conformational change occurs after the initial encounter complex is formed. The dissociation of the complex at equilibrium occurs in a single process of rate 0.095 s-1 at pH8.0 and 25 degrees C. Stopped-flow CD studies show that a slow decrease in ellipticity at 275 nm occurs with a rate of 1.3 s-1 when wild-type protein binds to kappa-chains, suggesting that the conformational transition might involve a change in environment around one or more tyrosine residues.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-103686,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-1436040,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-1618782,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-1698870,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-1909733,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-3881765,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-4156826,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-4628535,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-5013292,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-5086604,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-5723095,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-6234364,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-7051784,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-7608965,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-7716157,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-7947810,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-8010966,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-8280081,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-8461301,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-8493725,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-8654399,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-8767928,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-9054568,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10229674-9231896
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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 |
May
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pubmed:issn |
0264-6021
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
15
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pubmed:volume |
340 ( Pt 1)
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
193-9
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pubmed:dateRevised |
2011-7-28
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pubmed:meshHeading |
pubmed-meshheading:10229674-Amino Acid Substitution,
pubmed-meshheading:10229674-Bacterial Proteins,
pubmed-meshheading:10229674-Binding Sites,
pubmed-meshheading:10229674-Circular Dichroism,
pubmed-meshheading:10229674-Humans,
pubmed-meshheading:10229674-Hydrogen-Ion Concentration,
pubmed-meshheading:10229674-Immunoglobulin G,
pubmed-meshheading:10229674-Immunoglobulin kappa-Chains,
pubmed-meshheading:10229674-Kinetics,
pubmed-meshheading:10229674-Peptide Fragments,
pubmed-meshheading:10229674-Peptostreptococcus,
pubmed-meshheading:10229674-Protein Binding,
pubmed-meshheading:10229674-Protein Structure, Secondary,
pubmed-meshheading:10229674-Sequence Homology, Amino Acid,
pubmed-meshheading:10229674-Spectrometry, Fluorescence,
pubmed-meshheading:10229674-Static Electricity,
pubmed-meshheading:10229674-Temperature,
pubmed-meshheading:10229674-Thermodynamics,
pubmed-meshheading:10229674-Tryptophan,
pubmed-meshheading:10229674-Tyrosine
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pubmed:year |
1999
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pubmed:articleTitle |
Interactions between a single immunoglobulin-binding domain of protein L from Peptostreptococcus magnus and a human kappa light chain.
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pubmed:affiliation |
Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton, Hants. SO16 7PX, UK.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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