Overcoming hysteresis to attain reversible equilibrium folding for outer membrane phospholipase A in phospholipid bilayers.

ABSTRACT: The free energy of unfolding of a membrane protein from lipids into water (ΔG(o)(w,l)) describes its equilibrium thermodynamic stability. Knowing this parameter gives insight into a membrane protein's sequence-structure-energy relationships. However, there are few measures of membrane protein stability because of the technical difficulties associated with unfolded and partially folded states. Here, we describe the experimental process that allowed us to measure the ΔG(o)(w,l) of the outer membrane phospholipase A into large unilamellar vesicles (LUVs) of 1,2-dilauroyl-sn-glycero-3-phosphocholine. To arrive at this reversible folding condition, we screened a large number of experimental variables: temperature, incubation time, salt concentration, pH, lipid composition and liposome morpholog

SUBMITTER: Moon CP 

PROVIDER: S-EPMC3193555 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature