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Ligand‐induced formation of the leukotriene B 4 receptor‐G protein complex of human polymorphonuclear leukocytes
Author(s) -
Sherman Jeffrey W.,
Mendelson Michelle A.,
Boggs John M.,
Koo Catherine H.,
Goetzl Edward J.
Publication year - 1992
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240480405
Subject(s) - leukotriene b4 , ligand (biochemistry) , chemistry , receptor , leukotriene , microbiology and biotechnology , granulocyte , immunology , inflammation , biochemistry , biology , asthma
The components of the polymorphonuclear leukocyte (PMNL) receptor for leukotriene B 4 (LTB 4 ) were examined by Sephacryl S‐300 exclusion chromatography of PMNL membrane proteins, which were solubilized before and after the binding of [3H] LTB 4 . When the PMNL membranes were solubilized in 3‐[(3‐cholamidopropyl)‐dimethylammonio]‐1‐propanesulfonate (CHAPS) and filtered on Sephacryl S‐300 prior to addition of [3H] LTB 4 , the binding activity was associated with a 65 kD protein. In contrast, the radioactivity of [3H] LTB 4 bound to PMNL membranes prior to solubilization was recovered predominately with a 140 kD protein. When PMNL membranes had been pretreated with pertussis toxin, but not cholera toxin, before the addition of LTB 4 and subsequent solubilization, radioactivity was recovered predominately with the 65 kD protein. The addition of guanylylimidodiphosphate (GMP‐PNP), a nonhydrolyzable derivative of guanosine triphosphate (GTP), to PMNL membrane receptors bearing [3H] LTB 4 either prior to or after CHAPS solubilization reduced the yield of the 140 kD presumed LTB 4 receptor protein‐G protein complex. That the maximum specific binding of [35S] guanosine‐5′‐0‐3‐thiotriphosphate (GTP‐gammaS) to LTB 4 ‐binding proteins in the Sephacryl S‐300 effluent corresponded to the 140 kD protein supported the presence of a G protein in the LTB 4 receptor complex.