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PMID: 3541923 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Guanine nucleotide regulation of phospholipase C activity in permeabilized rabbit neutrophils. Inhibition by pertussis toxin and sensitization to submicromolar calcium concentrations.

The Biochemical journal ·Vol. 239 ·No. 1 ·1986-10-01 ·Pages 97-102

Bradford PG, Rubin RP

Abstract

Rabbit neutrophils labelled with [3H]inositol and permeabilized with saponin produced [3H]inositol trisphosphate (InsP3) when incubated with stable analogues of GTP or millimolar concentrations of Ca2+. [3H]InsP3 production elicited by guanosine 5'-[gamma-thio]triphosphate was enhanced by the chemoattractant formylmethionyl-leucyl-phenylalanine and inhibited by pertussis-toxin pretreatment. A pertussis-toxin-sensitive stimulation of [3H]InsP3 concentration was also observed with guanosine 5'-[beta gamma-imido]triphosphate, but not with guanosine 5'-[beta-thio]diphosphate or GTP. Millimolar Ca2+ alone was sufficient to stimulate [3H]InsP3 production; however, in the presence of guanosine 5'-[gamma-thio]triphosphate, the Ca2+ dose-response curve was shifted to submicromolar concentrations. These findings directly confirm the role of a pertussis-toxin-sensitive guanine nucleotide regulatory protein (G protein) in chemoattractant-stimulated phospholipase C activity in rabbit neutrophils. Moreover, the ability of guanine nucleotides to sensitize phospholipase C to physiologically relevant Ca2+ concentrations suggests that the role of the activated G protein may be to enhance the apparent affinity of phospholipase C for Ca2+ and thus to activate the enzyme without an increase in the Ca2+ concentration.

MeSH Terms
Adenosine Diphosphate/metabolism Animals Calcium/pharmacology Cell Membrane Permeability Guanine Nucleotides/pharmacology Guanosine 5'-O-(3-Thiotriphosphate) Guanosine Triphosphate/analogs & derivatives,pharmacology Inositol 1,4,5-Trisphosphate Inositol Phosphates/metabolism N-Formylmethionine Leucyl-Phenylalanine/pharmacology Neutrophils/drug effects,enzymology Pertussis Toxin Rabbits Thionucleotides/pharmacology Type C Phospholipases/metabolism Virulence Factors, Bordetella/pharmacology
Chemicals
Guanine Nucleotides Inositol Phosphates Thionucleotides Virulence Factors, Bordetella Guanosine 5'-O-(3-Thiotriphosphate) N-Formylmethionine Leucyl-Phenylalanine Adenosine Diphosphate Inositol 1,4,5-Trisphosphate Guanosine Triphosphate Pertussis Toxin Type C Phospholipases Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bradford P G
Rubin R P
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35 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2390097
Published
1986-10-01
Pages
97-102
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1147244
Subset
IM
Grants
NIADDK NIH HHS · AM-28029 · United States
NHLBI NIH HHS · HL-07357 · United States
External Links
PubMed source DOI: 10.1042/bj2390097 PMC Full Text View journal details
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