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Human substance P receptor expressed in Sf9 cells couples with multiple endogenous G proteins.

Publication ,  Journal Article
Nishimura, K; Frederick, J; Kwatra, MM
Published in: J Recept Signal Transduct Res
January 1998

To identify the G proteins involved in the function of human substance P receptor (hSPR), the receptor was expressed in Sf9 cells using the baculovirus expression system. Maximal hSPR expression was up to 65 pmol/mg membrane protein. The following data indicated that hSPR in Sf9 membranes is coupled to endogenous G proteins: 1) binding of agonist radioligand [125I]BHSP to the receptor was sensitive to guanine nucleotides; and 2) stimulation of the receptor increased [35S]GTPgammaS binding. The hSPR-associated G proteins were identified by photoaffinity labeling with [alpha-32P]-azidoanilido GTP ([alpha-32P]AAGTP), followed by immunoprecipitation of the labeled G proteins with antibodies specific for various Galpha-subunits. These experiments showed that stimulation of hSPR in Sf9 membranes activated multiple endogenous G proteins including Galpha(o), Galpha(q/11), and Galpha(s). While hSPR's ability to associate with Gq/11 is well-documented, the present study provides the first evidence of hSPR's potential to activate Galpha(o) and Galpha(s).

Duke Scholars

Published In

J Recept Signal Transduct Res

DOI

ISSN

1079-9893

Publication Date

January 1998

Volume

18

Issue

1

Start / End Page

51 / 65

Location

England

Related Subject Headings

  • Spodoptera
  • Recombinant Proteins
  • Receptors, Neurokinin-1
  • Membrane Proteins
  • Humans
  • GTP-Binding Proteins
  • Cell Line
  • Biochemistry & Molecular Biology
  • Baculoviridae
  • Animals
 

Citation

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Nishimura, K., Frederick, J., & Kwatra, M. M. (1998). Human substance P receptor expressed in Sf9 cells couples with multiple endogenous G proteins. J Recept Signal Transduct Res, 18(1), 51–65. https://doi.org/10.3109/10799899809039164
Nishimura, K., J. Frederick, and M. M. Kwatra. “Human substance P receptor expressed in Sf9 cells couples with multiple endogenous G proteins.J Recept Signal Transduct Res 18, no. 1 (January 1998): 51–65. https://doi.org/10.3109/10799899809039164.
Nishimura K, Frederick J, Kwatra MM. Human substance P receptor expressed in Sf9 cells couples with multiple endogenous G proteins. J Recept Signal Transduct Res. 1998 Jan;18(1):51–65.
Nishimura, K., et al. “Human substance P receptor expressed in Sf9 cells couples with multiple endogenous G proteins.J Recept Signal Transduct Res, vol. 18, no. 1, Jan. 1998, pp. 51–65. Pubmed, doi:10.3109/10799899809039164.
Nishimura K, Frederick J, Kwatra MM. Human substance P receptor expressed in Sf9 cells couples with multiple endogenous G proteins. J Recept Signal Transduct Res. 1998 Jan;18(1):51–65.
Journal cover image

Published In

J Recept Signal Transduct Res

DOI

ISSN

1079-9893

Publication Date

January 1998

Volume

18

Issue

1

Start / End Page

51 / 65

Location

England

Related Subject Headings

  • Spodoptera
  • Recombinant Proteins
  • Receptors, Neurokinin-1
  • Membrane Proteins
  • Humans
  • GTP-Binding Proteins
  • Cell Line
  • Biochemistry & Molecular Biology
  • Baculoviridae
  • Animals