Linked Life Data

10702266

Source:http://linkedlifedata.com/resource/pubmed/id/10702266

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2000-4-3
pubmed:abstractText
To determine the intracellular signaling mechanism of the 5-HT(2C) receptor endogenously expressed in choroid plexus epithelial cells, we implemented a strategy of targeted disruption of protein-protein interactions. This strategy entails the delivery of conjugated membrane-permeable peptides that disrupt domain interaction at specific steps in the signaling cascade. As proof of concept, two peptides targeted against receptor-G protein interaction domains were examined. Only G(q)CT, which targets the receptor-G(q) protein interacting domain, disrupted 5-HT(2C) receptor-mediated phosphatidylinositide hydrolysis. G(s)CT, targeting the receptor-G(s) protein, disrupted beta2 adrenergic receptor-mediated activation of cAMP but not 5-HT(2C) receptor-mediated phosphatidylinositide hydrolysis. The peptide MPS-PLCbeta1M, mimicking the domain of phospholipase Cbeta1 (PLCbeta1) interacting with active Galpha(q), also blocked 5-HT(2C) receptor activation. In contrast, peptides PLCbeta2M and Phos that bind to and sequester free Gbetagamma subunits were ineffective at blocking 5-HT(2C) receptor-mediated phosphoinositol turnover. However, both peptides disrupted Gbetagamma-mediated alpha(2A) adrenergic receptor activation of mitogen-activated protein kinase. These results provide the first direct demonstration that active Galpha(q) subunits mediate endogenous 5-HT(2C) receptor activation of PLCbeta and that Gbetagamma subunits released from Galpha(q) heterotrimeric proteins are not involved. Comparable results were obtained with metabotropic glutamate receptor 5 expressed in astrocytes. Thus, conjugated, membrane-permeable peptides are effective tools for the dissection of intracellular signals.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
10
pubmed:volume
275
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
7021-9
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:10702266-Amino Acid Sequence, pubmed-meshheading:10702266-Animals, pubmed-meshheading:10702266-Astrocytes, pubmed-meshheading:10702266-Cell Membrane Permeability, pubmed-meshheading:10702266-Cells, Cultured, pubmed-meshheading:10702266-Choroid Plexus, pubmed-meshheading:10702266-Epithelial Cells, pubmed-meshheading:10702266-GTP-Binding Proteins, pubmed-meshheading:10702266-Humans, pubmed-meshheading:10702266-Mitogen-Activated Protein Kinase Kinases, pubmed-meshheading:10702266-Molecular Sequence Data, pubmed-meshheading:10702266-Peptide Fragments, pubmed-meshheading:10702266-Rats, pubmed-meshheading:10702266-Rats, Sprague-Dawley, pubmed-meshheading:10702266-Receptor, Serotonin, 5-HT2C, pubmed-meshheading:10702266-Receptors, Serotonin, pubmed-meshheading:10702266-Type C Phospholipases, pubmed-meshheading:10702266-Virulence Factors, Bordetella
pubmed:year
2000
pubmed:articleTitle
Dissecting G protein-coupled receptor signaling pathways with membrane-permeable blocking peptides. Endogenous 5-HT(2C) receptors in choroid plexus epithelial cells.
pubmed:affiliation
Department of Pharmacology and Center for Molecular Neuroscience, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.