Physical determinants of β-barrel membrane protein folding in lipid vesicles.

Abstract:

:The spontaneous folding of two Neisseria outer membrane proteins, opacity-associated (Opa)(60) and Opa(50) into lipid vesicles was investigated by systematically varying bulk and membrane properties. Centrifugal fractionation coupled with sodium dodecyl sulfate polyacrylamide gel electrophoresis mobility assays enabled the discrimination of aggregate, unfolded membrane-associated, and folded membrane-inserted protein states as well as the influence of pH, ionic strength, membrane surface potential, lipid saturation, and urea on each. Protein aggregation was reduced with increasing lipid chain length, basic pH, low salt, the incorporation of negatively charged guest lipids, or by the addition of urea to the folding reaction. Insertion from the membrane-associated form was improved in shorter chain lipids, with more basic pH and low ionic strength; it is hindered by unsaturated or ether-linked lipids. The isolation of the physical determinants of insertion suggests that the membrane surface and dipole potentials are driving forces for outer membrane protein insertion and folding into lipid bilayers.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Dewald AH,Hodges JC,Columbus L

doi

10.1016/j.bpj.2011.03.025

subject

Has Abstract

pub_date

2011-05-04 00:00:00

pages

2131-40

issue

9

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(11)00373-0

journal_volume

100

pub_type

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