Rapid isolation of plasmalemma from cultured A431 cells: characterization of epidermal growth factor receptors.
A rapid method for the purification of plasma membrane from a relatively small number of A431 cells is described. The method is a simple, two-step differential centrifugation in the presence of Ca2+ that requires a total centrifugation time of 7 min. The membrane preparations contained a high level of epidermal growth factor (EGF) receptor activity demonstrated by both the quantity of specific ligand binding and the amount of EGF-dependent phosphorylation of the receptor and an exogenous substrate. EGF-dependent autophosphorylation identified the EGF receptor in the purified membranes as an undegraded 170-kDa protein.
Lin
P H
PH
University of California Experimental Pathology Group, Los Alamos National Laboratory, New Mexico 87545.
Selinfreund
R H
RH
Wakshull
E
E
Wharton
W
W
D000804:Angiotensin II / Q000032:analysis; D002462:Cell Membrane / Q000032:analysis*; D066246:ErbB Receptors / Q000032:analysis*; D006801:Humans; D010766:Phosphorylation; D014407:Tumor Cells, Cultured
D000804:Angiotensin II; D066246:ErbB Receptors