Linked Life Data

8089487

Source:http://linkedlifedata.com/resource/pubmed/id/8089487

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
7
pubmed:dateCreated
1994-10-14
pubmed:abstractText
The interactive residues for mouse mAb NC10.8, which binds a superpotent guanidinium sweetener N-(p-cyanophenyl)-N'-(diphenylmethyl)guanidinoacetic acid with high affinity (Kd = 5 nM), were examined by using radioligand competitive binding, photoaffinity labeling, absorption and fluorescence spectroscopy, computer-aided molecular modeling, and site-directed mutagenesis. Competitive ligand analogue binding data revealed important structural features and a pH sensitivity for ligand binding. Spectroscopy of the sweetener-mAb complex revealed ligand-induced fluorescence quenching and the presence of a charge-transfer band. Site-directed mutagenesis of L:96W abolished the ligand-induced fluorescence quenching and reduced Ab affinity. The apparent Kd increased from 5 nM to more than 200 nM after such modification. A theoretical model of the Fv region was generated with use of a knowledge-based algorithm, and this model was used to identify the locations of key residues in the complementarity determining regions. These experimental and theoretical studies support the prediction that the sweetener ligand coordinates with the following residues: L:34H contacts the cyanophenyl ring, L:27DR forms a salt bridge with the acetic acid moiety, L:96W forms a pi-pi interaction with the cyanophenyl ring, and H:95E contacts the positively charged aryl nitrogen. These studies are important to our understanding of Ab-ligand specificity and may also shed light on the important chemical motifs responsible for elevated levels of sweetness potency in organic compounds.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
AIM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0022-1767
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
153
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3059-69
pubmed:dateRevised
2011-11-17
pubmed:meshHeading
pubmed-meshheading:8089487-Acetic Acids, pubmed-meshheading:8089487-Amino Acid Sequence, pubmed-meshheading:8089487-Antibodies, Monoclonal, pubmed-meshheading:8089487-Base Sequence, pubmed-meshheading:8089487-Binding Sites, Antibody, pubmed-meshheading:8089487-Cloning, Molecular, pubmed-meshheading:8089487-Computer Simulation, pubmed-meshheading:8089487-DNA Primers, pubmed-meshheading:8089487-Guanidines, pubmed-meshheading:8089487-Ligands, pubmed-meshheading:8089487-Models, Molecular, pubmed-meshheading:8089487-Molecular Sequence Data, pubmed-meshheading:8089487-Mutagenesis, Site-Directed, pubmed-meshheading:8089487-Protein Structure, Tertiary, pubmed-meshheading:8089487-Spectrometry, Fluorescence, pubmed-meshheading:8089487-Structure-Activity Relationship, pubmed-meshheading:8089487-Sweetening Agents
pubmed:year
1994
pubmed:articleTitle
Identification of residues in monoclonal antibody NC10.8 that bind to the sweetener N-(p-cyanophenyl)-N'-(diphenylmethyl)guanidinoacetic acid by using radioligand binding, absorption and fluorescence spectroscopy, computer-aided molecular modeling, and site-directed mutagenesis.
pubmed:affiliation
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station 77843.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.