Synthesis and use of a novel N-formyl peptide derivative to isolate a human N-formyl peptide receptor cDNA

F Boulay, M Tardif, L Brouchon, P Vignais - Biochemical and biophysical …, 1990 - Elsevier
F Boulay, M Tardif, L Brouchon, P Vignais
Biochemical and biophysical research communications, 1990Elsevier
N-formyl-methionyl peptides are powerful chemoattractants which bind to specific receptors
on the neutrophil plasma membrane. A cDNA library from HL-60 cells, differentiated into
granulocytes highly responsive to N-formyl-methionyl peptides, was constructed in the COS
cell expression vector CDM8. A cDNA clone was isolated that conferred to COS cells the
ability to bind a new and highly efficient hydrophilic derivative of N-formyl-Met-Leu-Phe-Lys.
The transfected COS cells displayed two classes of binding sites with Kd values of 1–5nM …
Abstract
N-formyl-methionyl peptides are powerful chemoattractants which bind to specific receptors on the neutrophil plasma membrane. A cDNA library from HL-60 cells, differentiated into granulocytes highly responsive to N-formyl-methionyl peptides, was constructed in the COS cell expression vector CDM8. A cDNA clone was isolated that conferred to COS cells the ability to bind a new and highly efficient hydrophilic derivative of N-formyl-Met-Leu-Phe-Lys. The transfected COS cells displayed two classes of binding sites with Kd values of 1–5nM and 5–10 nM, respectively. The cDNA was 1.9 kb long with a 1050 bp open reading frame encoding a 350 residue protein. The hydropathy plot analysis revealed seven hydrophobic segments, a pattern quite similar to that of G protein-coupled receptors.
Elsevier