Sensitivity of orexin-A binding to phospholipase C inhibitors, neuropeptide Y, and secretin

J. K. Kane, H. Tanaka, S. L. Parker, Masashi Yanagisawa, M. D. Li

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The binding of [125I] orexin-A (Ox-A) to particulates from Chinese hamster ovary (CHO) cells expressing the cloned orexin-A receptor, or from rat forebrain areas, was sensitive to blockers of phosphatidylinositol-specific phospholipase C (PtdIns-PLC) U-73122 and ET-18-OCH3, little affected by phospholipase A2 inhibitor quinacrine, and not sensitive to D609, a xanthate inhibitor of phosphatidylcholine-selective PLC. Interaction of the receptor with a PtdIns-PLC was further indicated by a large sensitivity of the binding to Ca2+. Up to 50% of the binding was sensitive to the G-protein nucleotide site agonist GTP-γ-S. Ligand attachment to the orexin-A receptor thus depends on an association with both PtdIns-PLC and G-protein α-subunits. In all paradigms examined, the binding of [125I]orexin-A was competed by human/rat neuropeptide Y (hNPY) and porcine secretin with a potency similar to orexin-A (IC50 range 30-100 nM). The rank order of potency for NPY-related peptides was hNPY > porcine peptide YY (pPYY) > (Leu31, Pro34) human PYY > human PYY( 3-36) > hNPY free acid > human pancreatic polypeptide. Among secretin-related peptides, the rank order of potency was porcine secretin > orexin-A > human pituitary adenylate cyclase-activating peptide > orexin-B > porcine vasoactive intestinal peptide. Among opioid peptides, rat β-endorphin and camel δ-endorphin were much less active than NPY and secretin, and two enkephalins were inactive at 1 μM. In view of high abundance of NPY in forebrain, the above cross-reactivity could indicate a significant contribution of NPY to signaling via orexin-A receptors. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)959-965
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume272
Issue number3
DOIs
StatePublished - Jun 16 2000

Fingerprint

Secretin
Neuropeptide Y
Type C Phospholipases
Orexin Receptors
Rats
Phosphoinositide Phospholipase C
Endorphins
Swine
GTP-Binding Proteins
Peptides
Prosencephalon
Peptide YY
Quinacrine
Opioid Peptides
Enkephalins
Vasoactive Intestinal Peptide
Protein Subunits
Camelus
Programmable logic controllers
Guanosine Triphosphate

Keywords

  • Glucagon/secretin family
  • Neuropeptide cross-reactivity
  • Orexin-A receptor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Sensitivity of orexin-A binding to phospholipase C inhibitors, neuropeptide Y, and secretin. / Kane, J. K.; Tanaka, H.; Parker, S. L.; Yanagisawa, Masashi; Li, M. D.

In: Biochemical and Biophysical Research Communications, Vol. 272, No. 3, 16.06.2000, p. 959-965.

Research output: Contribution to journalArticle

@article{d589956bc76e4f91b1addf2e729838e7,
title = "Sensitivity of orexin-A binding to phospholipase C inhibitors, neuropeptide Y, and secretin",
abstract = "The binding of [125I] orexin-A (Ox-A) to particulates from Chinese hamster ovary (CHO) cells expressing the cloned orexin-A receptor, or from rat forebrain areas, was sensitive to blockers of phosphatidylinositol-specific phospholipase C (PtdIns-PLC) U-73122 and ET-18-OCH3, little affected by phospholipase A2 inhibitor quinacrine, and not sensitive to D609, a xanthate inhibitor of phosphatidylcholine-selective PLC. Interaction of the receptor with a PtdIns-PLC was further indicated by a large sensitivity of the binding to Ca2+. Up to 50{\%} of the binding was sensitive to the G-protein nucleotide site agonist GTP-γ-S. Ligand attachment to the orexin-A receptor thus depends on an association with both PtdIns-PLC and G-protein α-subunits. In all paradigms examined, the binding of [125I]orexin-A was competed by human/rat neuropeptide Y (hNPY) and porcine secretin with a potency similar to orexin-A (IC50 range 30-100 nM). The rank order of potency for NPY-related peptides was hNPY > porcine peptide YY (pPYY) > (Leu31, Pro34) human PYY > human PYY( 3-36) > hNPY free acid > human pancreatic polypeptide. Among secretin-related peptides, the rank order of potency was porcine secretin > orexin-A > human pituitary adenylate cyclase-activating peptide > orexin-B > porcine vasoactive intestinal peptide. Among opioid peptides, rat β-endorphin and camel δ-endorphin were much less active than NPY and secretin, and two enkephalins were inactive at 1 μM. In view of high abundance of NPY in forebrain, the above cross-reactivity could indicate a significant contribution of NPY to signaling via orexin-A receptors. (C) 2000 Academic Press.",
keywords = "Glucagon/secretin family, Neuropeptide cross-reactivity, Orexin-A receptor",
author = "Kane, {J. K.} and H. Tanaka and Parker, {S. L.} and Masashi Yanagisawa and Li, {M. D.}",
year = "2000",
month = "6",
day = "16",
doi = "10.1006/bbrc.2000.2880",
language = "English (US)",
volume = "272",
pages = "959--965",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Sensitivity of orexin-A binding to phospholipase C inhibitors, neuropeptide Y, and secretin

AU - Kane, J. K.

AU - Tanaka, H.

AU - Parker, S. L.

AU - Yanagisawa, Masashi

AU - Li, M. D.

PY - 2000/6/16

Y1 - 2000/6/16

N2 - The binding of [125I] orexin-A (Ox-A) to particulates from Chinese hamster ovary (CHO) cells expressing the cloned orexin-A receptor, or from rat forebrain areas, was sensitive to blockers of phosphatidylinositol-specific phospholipase C (PtdIns-PLC) U-73122 and ET-18-OCH3, little affected by phospholipase A2 inhibitor quinacrine, and not sensitive to D609, a xanthate inhibitor of phosphatidylcholine-selective PLC. Interaction of the receptor with a PtdIns-PLC was further indicated by a large sensitivity of the binding to Ca2+. Up to 50% of the binding was sensitive to the G-protein nucleotide site agonist GTP-γ-S. Ligand attachment to the orexin-A receptor thus depends on an association with both PtdIns-PLC and G-protein α-subunits. In all paradigms examined, the binding of [125I]orexin-A was competed by human/rat neuropeptide Y (hNPY) and porcine secretin with a potency similar to orexin-A (IC50 range 30-100 nM). The rank order of potency for NPY-related peptides was hNPY > porcine peptide YY (pPYY) > (Leu31, Pro34) human PYY > human PYY( 3-36) > hNPY free acid > human pancreatic polypeptide. Among secretin-related peptides, the rank order of potency was porcine secretin > orexin-A > human pituitary adenylate cyclase-activating peptide > orexin-B > porcine vasoactive intestinal peptide. Among opioid peptides, rat β-endorphin and camel δ-endorphin were much less active than NPY and secretin, and two enkephalins were inactive at 1 μM. In view of high abundance of NPY in forebrain, the above cross-reactivity could indicate a significant contribution of NPY to signaling via orexin-A receptors. (C) 2000 Academic Press.

AB - The binding of [125I] orexin-A (Ox-A) to particulates from Chinese hamster ovary (CHO) cells expressing the cloned orexin-A receptor, or from rat forebrain areas, was sensitive to blockers of phosphatidylinositol-specific phospholipase C (PtdIns-PLC) U-73122 and ET-18-OCH3, little affected by phospholipase A2 inhibitor quinacrine, and not sensitive to D609, a xanthate inhibitor of phosphatidylcholine-selective PLC. Interaction of the receptor with a PtdIns-PLC was further indicated by a large sensitivity of the binding to Ca2+. Up to 50% of the binding was sensitive to the G-protein nucleotide site agonist GTP-γ-S. Ligand attachment to the orexin-A receptor thus depends on an association with both PtdIns-PLC and G-protein α-subunits. In all paradigms examined, the binding of [125I]orexin-A was competed by human/rat neuropeptide Y (hNPY) and porcine secretin with a potency similar to orexin-A (IC50 range 30-100 nM). The rank order of potency for NPY-related peptides was hNPY > porcine peptide YY (pPYY) > (Leu31, Pro34) human PYY > human PYY( 3-36) > hNPY free acid > human pancreatic polypeptide. Among secretin-related peptides, the rank order of potency was porcine secretin > orexin-A > human pituitary adenylate cyclase-activating peptide > orexin-B > porcine vasoactive intestinal peptide. Among opioid peptides, rat β-endorphin and camel δ-endorphin were much less active than NPY and secretin, and two enkephalins were inactive at 1 μM. In view of high abundance of NPY in forebrain, the above cross-reactivity could indicate a significant contribution of NPY to signaling via orexin-A receptors. (C) 2000 Academic Press.

KW - Glucagon/secretin family

KW - Neuropeptide cross-reactivity

KW - Orexin-A receptor

UR - http://www.scopus.com/inward/record.url?scp=0034674247&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034674247&partnerID=8YFLogxK

U2 - 10.1006/bbrc.2000.2880

DO - 10.1006/bbrc.2000.2880

M3 - Article

C2 - 10860858

AN - SCOPUS:0034674247

VL - 272

SP - 959

EP - 965

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 3

ER -