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Related Experiment Videos

Guanine nucleotides decrease the affinity of substance P binding to its receptor.

J Luber-Narod1, N D Boyd, S E Leeman

  • 1Department of Physiology, University of Massachusetts Medical School, Worcester 01655.

European Journal of Pharmacology
|April 25, 1990
PubMed
Summary
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Guanine nucleotides significantly reduce substance P (SP) binding affinity to rat salivary gland membranes. This effect, caused by 5'-guanylyl imidodiphosphate (GppNHp), reverses upon nucleotide removal, confirming guanine nucleotide regulation.

Area of Science:

  • Biochemistry
  • Pharmacology
  • Neuroscience

Background:

  • Substance P (SP) is a neuropeptide involved in various physiological processes.
  • Guanine nucleotides are known to modulate receptor binding.
  • The interaction between guanine nucleotides and SP receptors requires further elucidation.

Purpose of the Study:

  • To investigate the inhibitory effect of guanine nucleotides on 125I-Bolton-Hunter conjugated substance P (125I-BHSP) binding to rat submaxillary gland membranes.
  • To quantify the changes in binding affinity and kinetics induced by guanine nucleotides.

Main Methods:

  • Radioligand binding assays using 125I-BHSP.
  • Scatchard analysis to determine binding affinity (Kd).
  • Determination of the concentration of SP required to inhibit half of the 125I-BHSP binding (IC50).

Related Experiment Videos

  • Measurement of the dissociation rate of 125I-BHSP in the presence of guanine nucleotides.

    • Main Results:

    • Guanine nucleotides, specifically 5 ext-guanylyl imidodiphosphate (GppNHp), markedly reduced the binding affinity of 125I-BHSP to rat submaxillary gland membranes.
    • Scatchard analysis revealed a greater than or equal to 20-fold increase in the dissociation constant (Kd).
    • The IC50 for SP increased approximately 30-fold, and the dissociation rate of 125I-BHSP increased about 100-fold upon GppNHp addition.
    • High-affinity binding was fully restored after the removal of the guanine nucleotide.

    Conclusions:

    • Guanine nucleotides induce a significant loss of binding affinity for substance P at its receptor.
    • The non-hydrolyzable guanine nucleotide analog GppNHp alters both the affinity and dissociation kinetics of SP binding.
    • These findings highlight the role of guanine nucleotides in regulating SP receptor function.