Linked Life Data

9554879

Source:http://linkedlifedata.com/resource/pubmed/id/9554879

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
1998-5-21
pubmed:abstractText
The molecular basis for recognition by human P2Y1 receptors of the novel, competitive antagonist 2'-deoxy-N6-methyladenosine 3', 5'-bisphosphate (MRS 2179) was probed using site-directed mutagenesis and molecular modeling. The potency of this antagonist was measured in mutant receptors in which key residues in the transmembrane helical domains (TMs) 3, 5, 6, and 7 were replaced by Ala or other amino acids. The capacity of MRS 2179 to block stimulation of phospholipase C promoted by 2-methylthioadenosine 5'-diphosphate (2-MeSADP) was lost in P2Y1 receptors having F226A, K280A, or Q307A mutations, indicating that these residues are critical for the binding of the antagonist molecule. Mutation of the residues His132, Thr222, and Tyr136 had an intermediate effect on the capacity of MRS 2179 to block the P2Y1 receptor. These positions therefore appear to have a modulatory role in recognition of this antagonist. F131A, H277A, T221A, R310K, or S317A mutant receptors exhibited an apparent affinity for MRS 2179 that was similar to that observed with the wild-type receptor. Thus, Phe131, Thr221, His277, and Ser317 are not essential for antagonist recognition. A computer-generated model of the human P2Y1 receptor was built and analyzed to help interpret these results. The model was derived through primary sequence comparison, secondary structure prediction, and three-dimensional homology building, using rhodopsin as a template, and was consistent with data obtained from mutagenesis studies. We have introduced a "cross-docking" procedure to obtain energetically refined 3D structures of the ligand-receptor complexes. Cross-docking simulates the reorganization of the native receptor structure induced by a ligand. A putative nucleotide binding site was localized and used to predict which residues are likely to be in proximity to agonists and antagonists. According to our model TM6 and TM7 are close to the adenine ring, TM3 and TM6 are close to the ribose moiety, and TM3, TM6, and TM7 are near the triphosphate chain.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0022-2623
pubmed:author
pubmed:issnType
Print
pubmed:day
23
pubmed:volume
41
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1456-66
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed-meshheading:9554879-Adenosine Diphosphate, pubmed-meshheading:9554879-Amino Acid Sequence, pubmed-meshheading:9554879-Animals, pubmed-meshheading:9554879-COS Cells, pubmed-meshheading:9554879-Enzyme Inhibitors, pubmed-meshheading:9554879-Enzyme-Linked Immunosorbent Assay, pubmed-meshheading:9554879-Humans, pubmed-meshheading:9554879-Ligands, pubmed-meshheading:9554879-Models, Molecular, pubmed-meshheading:9554879-Molecular Sequence Data, pubmed-meshheading:9554879-Mutagenesis, Site-Directed, pubmed-meshheading:9554879-Point Mutation, pubmed-meshheading:9554879-Protein Conformation, pubmed-meshheading:9554879-Protein Structure, Secondary, pubmed-meshheading:9554879-Purinergic P2 Receptor Agonists, pubmed-meshheading:9554879-Purinergic P2 Receptor Antagonists, pubmed-meshheading:9554879-Receptors, Purinergic P2, pubmed-meshheading:9554879-Receptors, Purinergic P2Y1, pubmed-meshheading:9554879-Type C Phospholipases
pubmed:year
1998
pubmed:articleTitle
Human P2Y1 receptor: molecular modeling and site-directed mutagenesis as tools to identify agonist and antagonist recognition sites.
pubmed:affiliation
Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0810, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't