Detecting protein-ligand binding on supported bilayers by local pH modulation.

ABSTRACT: Herein, we describe a highly sensitive technique for detecting protein-ligand binding at the liquid/solid interface. The method is based upon modulation of the interfacial pH when the protein binds. This change is detected by ortho-Texas Red DHPE, which is doped into supported phospholipid bilayers and used as a pH-sensitive dye. The dye molecule fluoresces strongly at acidic pH values but not basic ones and has an apparent pK(A) of 7.8 in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine membranes containing 0.5 mol % biotin-cap-PE. This method was used to detect antibiotin/biotin binding interactions as well as the binding of cholera toxin B subunits to GM(1). Since these proteins are negatively charged under the conditions of the experiment the interface became slightly more acidic upon

SUBMITTER: Jung H 

PROVIDER: S-EPMC3195364 | biostudies-literature | 2009 Jan

REPOSITORIES: biostudies-literature