Document Detail

Coupling of the human Y2 receptor for neuropeptide Y and peptide YY to guanine nucleotide inhibitory proteins in permeabilized SMS-KAN cells.
MedLine Citation:
PMID:  7830057     Owner:  NLM     Status:  MEDLINE    
Using guanine nucleotides, pertussis toxin, and specific antisera against the COOH-terminals of the alpha-subunits of Gi1/2, Gi3, and G(o), the binding and biological response of the Y2 receptor (Y2R) for peptide YY (PYY) was probed in SMS-KAN neuroblastoma cells. The specific binding of radiolabeled PYY exhibited a single apparent dissociation constant, KD = 76 pM for intact cells and KD = 906 pM for permeabilized cells. However, other data suggested existence of multiple receptor affinity states. A shift in KD and a decrease in apparent number of binding sites (Bmax) was observed in permeabilized cells when incubated with guanine nucleotides. By contrast, in membrane preparations guanine nucleotides induced only a decrease in Bmax. In intact cells, agonist exposure inhibited the intracellular accumulation of forskolin-stimulated cyclic AMP by 80% (IC50 = 420 nM) compared with 94% inhibition (IC50 = 380 nM) in permeabilized cells. In permeabilized cells, preincubation with antisera against alpha i1/2 and alpha i3 blocked the functional response of PYY, with anti-alpha i3 being the most potent; whereas anti-alpha o failed to affect the cyclic AMP levels. These results suggest that permeabilized SMS-KAN cells serve as a good model system for analysis of Y2R binding kinetics and functional response and that the Y2R interacts directly with several different GiS (but not G(o)).
C Freitag; A B Svendsen; N Feldthus; K Løssl; S P Sheikh
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  64     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  1995 Feb 
Date Detail:
Created Date:  1995-02-21     Completed Date:  1995-02-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  643-50     Citation Subset:  IM    
Department of Clinical Biochemistry, National University Hospital Rigshospitalet, Copenhagen, Denmark.
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MeSH Terms
Cell Membrane Permeability
Cyclic AMP / metabolism
GTP-Binding Proteins / immunology,  metabolism*
Guanine Nucleotides / pharmacology
Immune Sera / immunology
Intracellular Membranes / metabolism
Neuroblastoma / metabolism*,  pathology
Peptide YY
Peptides / pharmacology
Pertussis Toxin
Receptors, Gastrointestinal Hormone / metabolism*
Receptors, Neuropeptide Y / metabolism*
Tumor Cells, Cultured
Virulence Factors, Bordetella / pharmacology
Reg. No./Substance:
0/Guanine Nucleotides; 0/Immune Sera; 0/Peptides; 0/Receptors, Gastrointestinal Hormone; 0/Receptors, Neuropeptide Y; 0/Virulence Factors, Bordetella; 0/peptide YY receptor; 106388-42-5/Peptide YY; 60-92-4/Cyclic AMP; EC Toxin; EC 3.6.1.-/GTP-Binding Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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