Isolation and reconstitution of the N-formylpeptide receptor from HL-60 derived neutrophils.
A multifunctional receptor for N-formylpeptides exists on the membranes of neutrophils. This receptor has now been isolated from neutrophils derived from HL-60 promyelocytic leukemia cells. After solubilization by Nonidet-P40 and purification by affinity chromatography and HPLC the isolated receptor was reconstituted into egg phosphatidylcholine vesicles by SM-2 Bio-Bead removal of the Nonidet-P40. Analysis of the affinity and selectivity of the receptor was done by direct binding of two high-affinity ligands, formyl-Met-Leu-[3H]Phe-OH and formyl-Nle-Leu-Phe-[3H]Tyr-OH. The data suggest that the receptor can be isolated and reconstituted without apparent alteration of its binding affinity and selectivity, and that there appear to be no co-factors or subunits upon which these binding characteristics are dependent.
D002460:Cell Line; D006801:Humans; D007700:Kinetics; D015470:Leukemia, Myeloid, Acute; D008567:Membranes, Artificial; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D009504:Neutrophils / Q000737:chemistry*; D010713:Phosphatidylcholines; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000302:isolation & purification*; D044042:Receptors, Formyl Peptide; D012995:Solubility
D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D011956:Receptors, Cell Surface; D044042:Receptors, Formyl Peptide; D009240:N-Formylmethionine Leucyl-Phenylalanine