Isolation and reconstitution of the formylpeptide receptor from HL-60 derived neutrophils
Thesis/Dissertation
·
OSTI ID:7009629
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. SDS-PAGE of the affinity column eluate and the fraction obtained by HPLC showed one major band with an apparent molecular weight of 76,000. Analysis of the affinity and selectivity of the receptor was done by direct binding of two high-affinity ligands, formyl-Met-Leu({sup 3}H)Phe-OH and formyl-Nle-Leu-Phe({sup 3}H)Tyr-OH. The data suggest that the receptor can be purified to homogeneity and reconstituted without apparent alteration of its binding affinity and selectivity, and that there appear to be no co-factors of subunits upon which these binding characteristics are dependent.
- Research Organization:
- Virginia Commonwealth Univ., Richmond, VA (USA)
- OSTI ID:
- 7009629
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANIMAL CELLS
BIOLOGICAL MATERIALS
BIOLOGY
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
CYTOLOGY
FORMALDEHYDE
HYDROGEN COMPOUNDS
LEUKOCYTES
LIGANDS
LIQUID COLUMN CHROMATOGRAPHY
MATERIALS
MEMBRANE PROTEINS
MEMBRANES
MOLECULAR STRUCTURE
NEUTROPHILS
ORGANIC COMPOUNDS
PEPTIDES
PROTEINS
PURIFICATION
RECEPTORS
SEPARATION PROCESSES
TRITIUM COMPOUNDS
TUMOR CELLS
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
ANIMAL CELLS
BIOLOGICAL MATERIALS
BIOLOGY
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
CYTOLOGY
FORMALDEHYDE
HYDROGEN COMPOUNDS
LEUKOCYTES
LIGANDS
LIQUID COLUMN CHROMATOGRAPHY
MATERIALS
MEMBRANE PROTEINS
MEMBRANES
MOLECULAR STRUCTURE
NEUTROPHILS
ORGANIC COMPOUNDS
PEPTIDES
PROTEINS
PURIFICATION
RECEPTORS
SEPARATION PROCESSES
TRITIUM COMPOUNDS
TUMOR CELLS