Exploring the in meso crystallization mechanism by characterizing the lipid mesophase microenvironment during the growth of single transmembrane α-helical peptide crystals

Leonie T'Van Hag, Konstantin Knoblich, Shane A. Seabrook, Nigel M. Kirby, Stephen T. Mudie, Deborah Lau, Xu Li, Sally L. Gras, Xavier Mulet, Matthew E. Call, Melissa J. Call, Calum J. Drummond, Charlotte E. Conn

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13 Citations (Scopus)

Abstract

The proposed mechanism for in meso crystallization of transmembrane proteins suggests that a protein or peptide is initially uniformly dispersed in the lipid self-assembly cubic phase but that crystals grow from a local lamellar phase, which acts as a conduit between the crystal and the bulk cubic phase. However, there is very limited experimental evidence for this theory. We have developed protocols to investigate the lipid mesophase microenvironment during crystal growth using standard procedures readily available in crystallography laboratories. This technique was used to characterize the microenvironment during crystal growth of the DAP12-TM peptide using synchrotron small angle X-ray scattering (SAXS) with a micro-sized X-ray beam. Crystal growth was found to occur from the gyroid cubic mesophase. For one in four crystals, a highly oriented local lamellar phase was observed, providing supporting evidence for the proposed mechanism for in meso crystallization. A new observation of this study was that we can differentiate diffraction peaks from crystals grown in meso, from peaks originating from the surrounding lipid matrix, potentially opening up the possibility of high-throughput SAXS analysis of in meso grown crystals. This article is part of the themed issue 'Soft interfacial materials: from fundamentals to formulation'.

Original languageEnglish
Article number20150125
Number of pages17
JournalPhilosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences
Volume374
Issue number2072
DOIs
Publication statusPublished - 28 Jul 2016
Externally publishedYes

Keywords

  • Cubic mesophase
  • DAP12
  • In meso crystallization
  • Local lamellar phase

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