Isolation and reconstitution of the N‐formylpeptide receptor from HL‐60 derived neutrophils

FEBS Letters

Freer, Richard J.¹  Hoyle, Peter C.¹ 

[1] Department of Pharmacology and Toxicology, Medical College of Virginia, Richmond, VA 23298, USA

关键词: Receptor;    Solubilization;    Reconstitution;    Lipid vesicle;    Affinity chromatography;    Scatchard analysis;   

DOI  :  10.1016/0014-5793(84)80142-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown