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Proc Natl Acad Sci U S A. 2015 Aug 4;112(31):9632-7. doi: 10.1073/pnas.1501836112. Epub 2015 Jul 21.

Computational redesign of the lipid-facing surface of the outer membrane protein OmpA.

Author information

1
Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI 48824; Center for Advanced Biotechnology and Medicine and the Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ 08854;
2
Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI 48824; Department of Biosystems and Agricultural Engineering, Michigan State University, East Lansing, MI 48824.
3
Center for Advanced Biotechnology and Medicine and the Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ 08854; nanda@cabm.rutgers.edu.

Abstract

Advances in computational design methods have made possible extensive engineering of soluble proteins, but designed β-barrel membrane proteins await improvements in our understanding of the sequence determinants of folding and stability. A subset of the amino acid residues of membrane proteins interact with the cell membrane, and the design rules that govern this lipid-facing surface are poorly understood. We applied a residue-level depth potential for β-barrel membrane proteins to the complete redesign of the lipid-facing surface of Escherichia coli OmpA. Initial designs failed to fold correctly, but reversion of a small number of mutations indicated by backcross experiments yielded designs with substitutions to up to 60% of the surface that did support folding and membrane insertion.

KEYWORDS:

OmpA; membrane proteins; protein design; statistical potential; β-barrel

PMID:
26199411
PMCID:
PMC4534290
DOI:
10.1073/pnas.1501836112
[Indexed for MEDLINE]
Free PMC Article

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