Detergent solubilization of the EGF receptor from A431 cells.
Functional reconstitution of purified preparations of human epidermal growth factor receptor (EGFR) requires dissociation of the protein from its plasma membrane lipid environment.  Solubilization of membrane proteins in this manner requires the use of detergents, which are known to disrupt plasma membrane lipid/protein interactions.  We have investigated the ability of three nonionic detergents to solubilize the human EGFR selectively, and have also analyzed the effect of these various treatments on the intrinsic tyrosyl kinase activity of the receptor.  The nonionic detergent known as n-octyl glucoside (n-octyl beta-D-glucopyranoside) was found to give the best combination of selectivity, yield, and maintenance of enzymatic activity of the human EGFR.
Dayanidhi
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Division of Biology, NSCORT, Kansas State University, Manhattan 66506-4901, USA.
Rintoul
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DA
D002294:Carcinoma, Squamous Cell / Q000737:chemistry / Q000473:pathology / Q000648:ultrastructure; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure; D002478:Cells, Cultured; D003902:Detergents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D066246:ErbB Receptors / Q000032:analysis / Q000187:drug effects* / Q000502:physiology; D005960:Glucosides / Q000494:pharmacology*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000187:drug effects* / Q000502:physiology; D008567:Membranes, Artificial; D011092:Polyethylene Glycols / Q000494:pharmacology; D011489:Protein Denaturation; D014407:Tumor Cells, Cultured
D003902:Detergents; D005960:Glucosides; D008565:Membrane Proteins; D008567:Membranes, Artificial; C018619:octyl-beta-D-glucoside; D011092:Polyethylene Glycols; D066246:ErbB Receptors