Dissociation of the receptor for immunoglobulin E in mild detergents.
We previously showed that, in the absence of phospholipids, exposure of the tetrameric receptor for immunoglobulin E to mild detergents dissociates the intact beta chain and two gamma chains from the alpha chains. Having developed a practical method for assaying the dissociation, we have now explored a variety of different detergents, detergent concentrations, temperatures, times, salts, pHs, and other factors that influence the detergent-induced dissociation. Our findings should be useful for optimizing the stability of the receptor and for future studies on recombination of the subunits. The data suggest the following: (1) The critical perturbant is micellar detergent. (2) Unlike solubilization of membranes, where a molar ratio of micellar detergent:lipid of 2 is adequate, dissociation of the receptor is incomplete even at molar ratios of micellar detergent:receptor of greater than 10(5) and may be limited by a reversible component. (3) Detergents that are best for solubilizing membranes are also best for dissociating the receptors. (4) The latter observation and other data implicate bound lipid as stabilizing the receptor. Our findings may be applicable to the study of interactions between membrane proteins in general.
Kinet
J P
JP
Alcaraz
G
G
Leonard
A
A
Wank
S
S
Metzger
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D001491:Basophils / Q000276:immunology; D002460:Cell Line; D003902:Detergents; D007073:Immunoglobulin E / Q000378:metabolism*; D007700:Kinetics; D007942:Leukemia, Experimental / Q000276:immunology; D046911:Macromolecular Substances; D008823:Micelles; D011092:Polyethylene Glycols; D051381:Rats; D011961:Receptors, Fc / Q000302:isolation & purification / Q000378:metabolism*; D017455:Receptors, IgE; D011971:Receptors, Immunologic / Q000302:isolation & purification / Q000378:metabolism*; D012995:Solubility; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D000911:Antibodies, Monoclonal; D003902:Detergents; D046911:Macromolecular Substances; D008823:Micelles; D011961:Receptors, Fc; D017455:Receptors, IgE; D011971:Receptors, Immunologic; D007073:Immunoglobulin E; D011092:Polyethylene Glycols