Journal article

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

Leonie van 't 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

PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY A-MATHEMATICAL PHYSICAL AND ENGINEERING SCIENCES | ROYAL SOC | Published : 2016

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 sc..

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Grants

Awarded by National Health and Medical Research Council (NHMRC) of Australia


Awarded by Victorian Government (VESKI Innovation Fellowship)


Awarded by QEII Fellowship from the Australian Research Council (ARC)


Awarded by ARC Future Fellowship


Awarded by ARC Dairy Innovation Hub


Funding Acknowledgements

This work was supported by grants from the National Health and Medical Research Council (NHMRC) of Australia (GNT1011352 to M.J.C. and M.E.C.; Independent Research Institutes Infrastructure Support Scheme (IRIISS) to the Walter and Eliza Hall Institute of Medical Research (WEHI)) and the Victorian Government (VESKI Innovation Fellowship VIF12 to M.E.C.; Operational Infrastructure Support to WEHI). M.E.C. is supported by a QEII Fellowship (DP110104369) from the Australian Research Council (ARC). M.J.C. is supported by an ARC Future Fellowship (FT120100145). S.L.G. is supported by the ARC Dairy Innovation Hub IH120100005.