Identification and functional studies of a specific peptide YY-preferring receptor in dog adipocytes.
Specific binding sites for peptide YY (PYY) and neuropeptide Y (NPY) as well as functional responses were identified in dog adipocytes. Studies were carried out using the radioligand [125I-Tyr1]monoiodo-PYY on crude adipocyte membranes. [125I]PYY bound to dog adipocyte membranes with a high affinity (156 +/- 24 pM) and binding capacity of 314 +/- 48 fmol/mg protein. Competition studies revealed a higher affinity of the binding sites for PYY than NPY (inhibition constants were 118 +/- 17 pM and 300 +/- 53 pM, respectively, P < or = 0.001). NPY analogs displaced [125I]PYY specific binding with the following order of potency: NPY-(13-36) > NPY-(18-36) > NPY-(22-36) >> [Leu31-Pro34]NPY. Neither adrenergic nor adenosine agents (activating or inhibiting other antilipolytic systems) interacted with [125I]PYY binding sites. So [125I]PYY binding was specific, saturable, and reversible. Lipolysis experiments performed with PYY, NPY, and NPY analogs confirm the relative order of potency found in competition experiments. The data agree with the definition of PYY-preferring receptor which resembles a Y2 receptor subtype since NPY-(13-36), a specific Y2 receptor agonist, inhibited binding and lipolysis in a similar way to PYY, whereas [Leu31-Pro34]NPY did not. No difference was observed in the antilipolytic response between IC50 values measured on omental, perirenal, and subcutaneous fat deposits. Moreover, PYY and NPY (10(-6) M) significantly attenuated forskolin-stimulated cAMP levels, involving inhibition of adenylyl cyclase as a transmembrane signaling mechanism. Cross-linking of bound [125I]PYY to membranes indicated that the mol wt of the receptor was 62K. The relative importance of such a receptor on fat cells alongside another powerful antilipolytic receptor--the alpha 2-adrenoceptor--is discussed.
Castan
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I
Institut National de la Santé et de la Recherche Médicale, INSERM U317, Institut L. Bugnard, CHU Rangueil, Toulouse, France.
Valet
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P
Voisin
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T
Quideau
N
N
Laburthe
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M
Lafontan
M
M
D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D004285:Dogs; D005768:Gastrointestinal Hormones / Q000378:metabolism; D008066:Lipolysis; D008970:Molecular Weight; D009478:Neuropeptide Y / Q000378:metabolism / Q000494:pharmacology; D019894:Peptide YY; D010455:Peptides / Q000378:metabolism* / Q000494:pharmacology; D011869:Radioligand Assay; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D017476:Receptors, Neuropeptide Y / Q000378:metabolism
D005768:Gastrointestinal Hormones; D009478:Neuropeptide Y; D010455:Peptides; D011956:Receptors, Cell Surface; D017476:Receptors, Neuropeptide Y; D019894:Peptide YY