VIP activates G(s) and G(i3) in rat alveolar macrophages and G(s) in HEK293 cells transfected with the human VPAC1 receptor

S. M. Shreeve; S. P. Sreedharan; M. P. Hacker; D. E. Gannon; M. J. Morgan

doi:10.1006/bbrc.2000.2879

VIP activates G(s) and G(i3) in rat alveolar macrophages and G(s) in HEK293 cells transfected with the human VPAC₁ receptor

S. M. Shreeve, S. P. Sreedharan, M. P. Hacker, D. E. Gannon, M. J. Morgan

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

We have characterized vasoactive intestinal peptide (VIP) receptor/G-protein coupling in rat alveolar macrophage (AM) membranes and find that pertussis toxin treatment and antisera against G(αi3) and G(αs) reduce high-affinity ¹²⁵I-VIP binding, indicating that both G(αs) and G(αi3) couple to the VIP-receptor. The predominant VIP-receptor subtype in AM is VPAC₁ and we examined the G-protein interactions of the human VPAC1 that had been transfected into HEK293 cells. VPAC1 has a molecular mass of 56 kDa; GTP analogs reduced ¹²⁵I-VIP binding to this protein demonstrating that high-affinity binding of VIP to the receptor requires coupling to G-protein. Functional VIP/VPAC₁/ G-protein complexes were captured by covalent cross-linking and analyzed by Western blotting. The transfected human VPAC1 receptor in HEK293 was found to be coupled to G(αs) but not G(αi) or G(αq). Furthermore, pertussis toxin treatment had no effect on VPAC₁/G-protein coupling in these cells. These observations suggest that the G-proteins activated by VPAC1 may be dependent upon species and cell type. (C) 2000 Academic Press.

Original language	English (US)
Pages (from-to)	922-928
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	272
Issue number	3
DOIs	https://doi.org/10.1006/bbrc.2000.2879
State	Published - Jun 16 2000
Externally published	Yes

Keywords

G(i3)
G-protein
G-protein, inhibitory
Human VPAC
Lung
Rat alveolar macrophages
Receptor signaling
Vasoactive intestinal peptide
Vasoactive intestinal peptide receptors

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1006/bbrc.2000.2879

Cite this

@article{40ed8d4432534af69f64a2acdfcdb877,

title = "VIP activates G(s) and G(i3) in rat alveolar macrophages and G(s) in HEK293 cells transfected with the human VPAC1 receptor",

abstract = "We have characterized vasoactive intestinal peptide (VIP) receptor/G-protein coupling in rat alveolar macrophage (AM) membranes and find that pertussis toxin treatment and antisera against G(αi3) and G(αs) reduce high-affinity 125I-VIP binding, indicating that both G(αs) and G(αi3) couple to the VIP-receptor. The predominant VIP-receptor subtype in AM is VPAC1 and we examined the G-protein interactions of the human VPAC1 that had been transfected into HEK293 cells. VPAC1 has a molecular mass of 56 kDa; GTP analogs reduced 125I-VIP binding to this protein demonstrating that high-affinity binding of VIP to the receptor requires coupling to G-protein. Functional VIP/VPAC1/ G-protein complexes were captured by covalent cross-linking and analyzed by Western blotting. The transfected human VPAC1 receptor in HEK293 was found to be coupled to G(αs) but not G(αi) or G(αq). Furthermore, pertussis toxin treatment had no effect on VPAC1/G-protein coupling in these cells. These observations suggest that the G-proteins activated by VPAC1 may be dependent upon species and cell type. (C) 2000 Academic Press.",

keywords = "G(i3), G-protein, G-protein, inhibitory, Human VPAC, Lung, Rat alveolar macrophages, Receptor signaling, Vasoactive intestinal peptide, Vasoactive intestinal peptide receptors",

author = "Shreeve, {S. M.} and Sreedharan, {S. P.} and Hacker, {M. P.} and Gannon, {D. E.} and Morgan, {M. J.}",

note = "Funding Information: The authors thank Arun Basu and Mary Gundell for their excellent technical assistance. This work was supported by the Vermont Chapter of the American Heart Association awarded to S.M.S.",

year = "2000",

month = jun,

day = "16",

doi = "10.1006/bbrc.2000.2879",

language = "English (US)",

volume = "272",

pages = "922--928",

journal = "Biochemical and Biophysical Research Communications",

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TY - JOUR

T1 - VIP activates G(s) and G(i3) in rat alveolar macrophages and G(s) in HEK293 cells transfected with the human VPAC1 receptor

AU - Shreeve, S. M.

AU - Sreedharan, S. P.

AU - Hacker, M. P.

AU - Gannon, D. E.

AU - Morgan, M. J.

N1 - Funding Information: The authors thank Arun Basu and Mary Gundell for their excellent technical assistance. This work was supported by the Vermont Chapter of the American Heart Association awarded to S.M.S.

PY - 2000/6/16

Y1 - 2000/6/16

N2 - We have characterized vasoactive intestinal peptide (VIP) receptor/G-protein coupling in rat alveolar macrophage (AM) membranes and find that pertussis toxin treatment and antisera against G(αi3) and G(αs) reduce high-affinity 125I-VIP binding, indicating that both G(αs) and G(αi3) couple to the VIP-receptor. The predominant VIP-receptor subtype in AM is VPAC1 and we examined the G-protein interactions of the human VPAC1 that had been transfected into HEK293 cells. VPAC1 has a molecular mass of 56 kDa; GTP analogs reduced 125I-VIP binding to this protein demonstrating that high-affinity binding of VIP to the receptor requires coupling to G-protein. Functional VIP/VPAC1/ G-protein complexes were captured by covalent cross-linking and analyzed by Western blotting. The transfected human VPAC1 receptor in HEK293 was found to be coupled to G(αs) but not G(αi) or G(αq). Furthermore, pertussis toxin treatment had no effect on VPAC1/G-protein coupling in these cells. These observations suggest that the G-proteins activated by VPAC1 may be dependent upon species and cell type. (C) 2000 Academic Press.

AB - We have characterized vasoactive intestinal peptide (VIP) receptor/G-protein coupling in rat alveolar macrophage (AM) membranes and find that pertussis toxin treatment and antisera against G(αi3) and G(αs) reduce high-affinity 125I-VIP binding, indicating that both G(αs) and G(αi3) couple to the VIP-receptor. The predominant VIP-receptor subtype in AM is VPAC1 and we examined the G-protein interactions of the human VPAC1 that had been transfected into HEK293 cells. VPAC1 has a molecular mass of 56 kDa; GTP analogs reduced 125I-VIP binding to this protein demonstrating that high-affinity binding of VIP to the receptor requires coupling to G-protein. Functional VIP/VPAC1/ G-protein complexes were captured by covalent cross-linking and analyzed by Western blotting. The transfected human VPAC1 receptor in HEK293 was found to be coupled to G(αs) but not G(αi) or G(αq). Furthermore, pertussis toxin treatment had no effect on VPAC1/G-protein coupling in these cells. These observations suggest that the G-proteins activated by VPAC1 may be dependent upon species and cell type. (C) 2000 Academic Press.

KW - G(i3)

KW - G-protein

KW - G-protein, inhibitory

KW - Human VPAC

KW - Lung

KW - Rat alveolar macrophages

KW - Receptor signaling

KW - Vasoactive intestinal peptide

KW - Vasoactive intestinal peptide receptors

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U2 - 10.1006/bbrc.2000.2879

DO - 10.1006/bbrc.2000.2879

M3 - Article

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AN - SCOPUS:0034674273

SN - 0006-291X

VL - 272

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JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

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