Method for measuring the unbinding energy of strongly-bound membrane-associated proteins.

ABSTRACT: We describe a new method to measure the activation energy for unbinding (enthalpy ΔH*_u and free energy ΔG*_u) of a strongly-bound membrane-associated protein from a lipid membrane. It is based on measuring the rate of release of a liposome-bound protein during centrifugation on a sucrose gradient as a function of time and temperature. The method is used to determine ΔH*_u and ΔG*_u for the soluble dengue virus envelope protein (sE) strongly bound to 80:20 POPC:POPG liposomes at pH5.5. ΔH*_u is determined from the Arrhenius equation whereas ΔG*_u is determined by fitting the data to a model based on mean first passage time for escape from a potential well. The binding free energy ΔG_b of sE was also measured at the same

SUBMITTER: Bauve E 

PROVIDER: S-EPMC5266593 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature