Molecular cloning of the fMet-Leu-Phe receptor from neutrophils

Kathleen M. Thomas, Hae Yung Pyun, Javier Navarro

Research output: Contribution to journalArticle

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Abstract

The bacterial chemotactic peptide fMet-Leu-Phe (fMLP) activates neutrophils upon binding to surface receptors. In a previous communication we reported the functional reconstitution of the fMLP receptor in Xenopus laevis oocytes (Coats, W. D., and Navarro, J. (1990) J. Biol. Chem. 265, 5964-5966). In this work we report the isolation of the cDNA encoding the fMLP receptor from neutrophils. A rabbit neutrophil cDNA library was screened with an oligonucleotide probe deduced from the nucleotide sequence of G-protein-coupled receptors, and a cDNA encoding the fMLP receptor was isolated. This cDNA was characterized according to the following criteria: 1) Analysis of the deduced amino acid sequence revealed that the clone belongs to a G-protein-coupled receptor. 2) Tissue distribution analysis of the mRNA indicated that the message is only found in neutrophils. 3) In vitro translation of the message revealed a protein corresponding in size to the deglycosylated fMLP receptor. 4) X. laevis oocytes injected with the fMLP receptor message exhibited fMLP-dependent calcium mobilization and specific binding to the fMLP analog 125I-labeled fNle-Leu-Phe-Nle-Tyr-Lys (where NIe is norleucine and fNle is formylnorleucine). The molecular cloning of the fMLP receptor should provide the framework to analyze the relationship between structure, function, and regulation of this receptor.

Original languageEnglish (US)
Pages (from-to)20061-20064
Number of pages4
JournalJournal of Biological Chemistry
Volume265
Issue number33
StatePublished - Nov 25 1990
Externally publishedYes

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Formyl Peptide Receptor
Cloning
Molecular Cloning
Neutrophils
N-Formylmethionine Leucyl-Phenylalanine
Complementary DNA
Xenopus laevis
G-Protein-Coupled Receptors
Oocytes
Norleucine
Oligonucleotide Probes
Protein Sequence Analysis
Tissue Distribution
Gene Library
Nucleotides
Clone Cells
Communication
Tissue
Rabbits
Calcium

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular cloning of the fMet-Leu-Phe receptor from neutrophils. / Thomas, Kathleen M.; Pyun, Hae Yung; Navarro, Javier.

In: Journal of Biological Chemistry, Vol. 265, No. 33, 25.11.1990, p. 20061-20064.

Research output: Contribution to journalArticle

Thomas, Kathleen M. ; Pyun, Hae Yung ; Navarro, Javier. / Molecular cloning of the fMet-Leu-Phe receptor from neutrophils. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 33. pp. 20061-20064.
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