A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes
Citation:
Caffrey, M., A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes, Acta Crystallographica Section F Structural Biology Communications, 71, 2015, 3 - 18Download Item:
Abstract:
The lipid cubic phase or
in meso
method is a robust approach for crystallizing
membrane proteins for structure determination. The uptake of the method is
such that it is experiencing what can only be described as explosive growth. This
timely, comprehensive and up-to-date review introduces the reader to the
practice of
in meso
crystallogenesis, to the associated challenges and to their
solutions. A model of how crystallization comes about mechanistically is
presented for a more rational approach to crystallization. The possible
involvement of the lamellar and inverted hexagonal phases in crystallogenesis
and the application of the method to water-soluble, monotopic and lipid-
anchored proteins are addressed. How to set up trials manually and
automatically with a robot is introduced with reference to open-access online
videos that provide a practical guide to all aspects of the method. These range
from protein reconstitution to crystal harvesting from the hosting mesophase,
which is noted for its viscosity and stickiness. The sponge phase, as an alternative
medium in which to perform crystallization, is described. The compatibility of
the method with additive lipids, detergents, precipitant-screen components and
materials carried along with the protein such as denaturants and reducing agents
is considered. The powerful host and additive lipid-screening strategies are
described along with how samples that have low protein concentration and
cell-free expressed protein can be used. Assaying the protein reconstituted in
the bilayer of the cubic phase for function is an important element of quality
control and is detailed. Host lipid design for crystallization at low temperatures
and for large proteins and complexes is outlined. Experimental phasing by
heavy-atom derivatization, soaking or co-crystallization is routine and the
approaches that have been implemented to date are described. An overview and
a breakdown by family and function of the close to 200 published structures that
have been obtained using
in meso
-grown crystals are given. Recommendations
for conducting the screening process to give a more productive outcome are
summarized. The fact that the
in meso
method also works with soluble proteins
should not be overlooked. Recent applications of the method for
in situ
serial
crystallography at X-ray free-electron lasers and synchrotrons are described.
The review ends with a view to the future and to the bright prospects for the
method, which continues to contribute to our understanding of the molecular
mechanisms of some of nature’s most valued proteinaceous robots.
Sponsor
Grant Number
Science Foundation Ireland (SFI)
12/IA/1255
OPSONA Therapeutics LTD under the Innovations Partnership Program
Author's Homepage:
http://people.tcd.ie/mcaffreDescription:
PUBLISHED
Author: CAFFREY, MARTIN
Sponsor:
Science Foundation Ireland (SFI)OPSONA Therapeutics LTD under the Innovations Partnership Program
Type of material:
Journal ArticleCollections
Series/Report no:
Acta Crystallographica Section F Structural Biology Communications71
Availability:
Full text availableSubject:
X-ray free-electron lasers, lipid cubic phaseDOI:
http://dx.doi.org/10.1107/S2053230X14026843Metadata
Show full item recordThe following license files are associated with this item: