Monitoring of detergent binding to amphiphilic proteins by means of micelles containing the hydrophobic dye Sudan Black B.
A simple method for detecting micellar binding of Triton X-100 to amphiphilic proteins is described. The hydrophobic dye Sudan Black B is incorporated into Triton micelles. Binding of the coloured micelles to serum apoliproteins, as well as to amphiphilic proteins, or erythrocyte and fat globule membranes renders these visible as dark bands after sucrose density gradient centrifugation. In contrast, the hydrophilic proteins present in lipoprotein-free serum do not show detergent binding. The method does not permit accurate quantification of detergent binding, but may serve as a pilot procedure for initial detection of amphiphilic proteins and for monitoring their isolation from crude solubilized membrane material. The sensitivity of the assay corresponds to that obtained with [3H]Triton X-100.
D000818:Animals; D001391:Azo Compounds; D001798:Blood Proteins; D002417:Cattle; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D004912:Erythrocytes; D006801:Humans; D008565:Membrane Proteins / Q000097:blood; D008823:Micelles; D008894:Milk Proteins; D009281:Naphthalenes; D017830:Octoxynol; D011092:Polyethylene Glycols; D011485:Protein Binding; D011506:Proteins
D001391:Azo Compounds; D001798:Blood Proteins; D008565:Membrane Proteins; D008823:Micelles; D008894:Milk Proteins; D009281:Naphthalenes; D011506:Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C016118:Sudan Black B