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Title: Membrane Protein Crystallization in Lipidic Mesophases. Hosting Lipid Effects on the Crystallization and Structure of a Transmembrane Peptide

Abstract

Gramicidin is an apolar pentadecapeptide antibiotic consisting of alternating d- and l-amino acids. It functions, in part, by creating pores in membranes of susceptible cells rendering them leaky to monovalent cations. The peptide should be able to traverse the host membrane either as a double-stranded, intertwined double helix (DSDH) or as a head-to-head single-stranded helix (HHSH). Current structure models are based on macromolecular X-ray crystallography (MX) and nuclear magnetic resonance (NMR). However, the HHSH form has only been observed by NMR. The shape and size of the different gramicidin conformations differ. We speculated therefore that reconstituting it into a lipidic mesophase with bilayers of different microstructures would preferentially stabilize one form over the other. By using such mesophases for in meso crystallogenesis, the expectation was that at least one would generate crystals of gramicidin in the HHSH form for structure determination by MX. This was tested using commercial and in-house synthesized lipids that support in meso crystallogenesis. Lipid acyl chain lengths were varied from 14 to 18 carbons to provide mesophases with a range of bilayer thicknesses. Unexpectedly, all lipids produced high-quality, structure-grade crystals with gramicidin only in the DSDH conformation.

Authors:
; ; ;  [1]
  1. (Trinity)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
FOREIGNNIH
OSTI Identifier:
1023655
Resource Type:
Journal Article
Journal Name:
Cryst. Growth Des.
Additional Journal Information:
Journal Volume: 11; Journal Issue: (4) ; 04, 2011; Journal ID: ISSN 1528-7483
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; ANTIBIOTICS; CATIONS; CRYSTALLIZATION; CRYSTALLOGRAPHY; CRYSTALS; HOST; LIPIDS; MEMBRANE PROTEINS; MEMBRANES; NUCLEAR MAGNETIC RESONANCE; PEPTIDES; RANGE; SHAPE; SIZE

Citation Formats

Hfer, Nicole, Aragao, David, Lyons, Joseph A., and Caffrey, Martin. Membrane Protein Crystallization in Lipidic Mesophases. Hosting Lipid Effects on the Crystallization and Structure of a Transmembrane Peptide. United States: N. p., 2011. Web. doi:10.1021/cg101384p.
Hfer, Nicole, Aragao, David, Lyons, Joseph A., & Caffrey, Martin. Membrane Protein Crystallization in Lipidic Mesophases. Hosting Lipid Effects on the Crystallization and Structure of a Transmembrane Peptide. United States. doi:10.1021/cg101384p.
Hfer, Nicole, Aragao, David, Lyons, Joseph A., and Caffrey, Martin. Wed . "Membrane Protein Crystallization in Lipidic Mesophases. Hosting Lipid Effects on the Crystallization and Structure of a Transmembrane Peptide". United States. doi:10.1021/cg101384p.
@article{osti_1023655,
title = {Membrane Protein Crystallization in Lipidic Mesophases. Hosting Lipid Effects on the Crystallization and Structure of a Transmembrane Peptide},
author = {Hfer, Nicole and Aragao, David and Lyons, Joseph A. and Caffrey, Martin},
abstractNote = {Gramicidin is an apolar pentadecapeptide antibiotic consisting of alternating d- and l-amino acids. It functions, in part, by creating pores in membranes of susceptible cells rendering them leaky to monovalent cations. The peptide should be able to traverse the host membrane either as a double-stranded, intertwined double helix (DSDH) or as a head-to-head single-stranded helix (HHSH). Current structure models are based on macromolecular X-ray crystallography (MX) and nuclear magnetic resonance (NMR). However, the HHSH form has only been observed by NMR. The shape and size of the different gramicidin conformations differ. We speculated therefore that reconstituting it into a lipidic mesophase with bilayers of different microstructures would preferentially stabilize one form over the other. By using such mesophases for in meso crystallogenesis, the expectation was that at least one would generate crystals of gramicidin in the HHSH form for structure determination by MX. This was tested using commercial and in-house synthesized lipids that support in meso crystallogenesis. Lipid acyl chain lengths were varied from 14 to 18 carbons to provide mesophases with a range of bilayer thicknesses. Unexpectedly, all lipids produced high-quality, structure-grade crystals with gramicidin only in the DSDH conformation.},
doi = {10.1021/cg101384p},
journal = {Cryst. Growth Des.},
issn = {1528-7483},
number = (4) ; 04, 2011,
volume = 11,
place = {United States},
year = {2011},
month = {9}
}