Detergent solubilization of human neutrophil leukotriene B4 receptors.
Specific leukotriene B4 (LTB4) receptors in human neutrophils were solubilized by treatment of "receptor fraction" membranes with the zwitterionic detergent (3-[(3-cholamidopropyl)-dimethylammonio]1-propane sulfonate (CHAPS). The soluble receptors were assayed by polyethylene glycol (PEG) precipitation coupled with Millipore filtration. The solubilized receptors retained all of the characteristics of the receptor sites in intact neutrophils. The binding of LTB4 was rapid, reversible and stereospecific. Mathematical modeling analysis revealed biphasic binding of [3H] LTB4 indicating two classes of binding sites. The high affinity binding site had a dissociation constant of 1.93 nM and Bmax of 281 fmoles/mg protein; the low affinity binding site had a dissociation constant of 78.92 nM and Bmax of 2522 fmoles/mg protein. Competitive binding experiments with structural analogs of LTB4 demonstrate that the interaction between LTB4 and its binding site is stereospecific and correlates with the relative biological activity of the analogs. These data suggest that it may be possible to purify the LTB4 receptor from human neutrophil membranes.
Lin
A H
AH
Ruppel
P L
PL
Gorman
R R
RR
D002793:Cholic Acids; D006801:Humans; D007700:Kinetics; D007975:Leukotriene B4 / Q000378:metabolism; D009504:Neutrophils / Q000032:analysis*; D011971:Receptors, Immunologic / Q000097:blood* / Q000302:isolation & purification; D018102:Receptors, Leukotriene B4; D012995:Solubility; D013329:Structure-Activity Relationship; D013997:Time Factors
D002793:Cholic Acids; D011971:Receptors, Immunologic; D018102:Receptors, Leukotriene B4; D007975:Leukotriene B4; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate