Membrane-protein crystallization in cubo: Temperature-dependent phase behaviour of monoolein-detergent mixtures

Sennoga, Charles, Heron, Andrew, Seddon, John M., Templer, Richard H. and Hankamer, Ben (2003) Membrane-protein crystallization in cubo: Temperature-dependent phase behaviour of monoolein-detergent mixtures. Acta Crystallographica Section D: Biological Crystallography, 59 2: 239-246. doi:10.1107/S0907444902020772

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Author Sennoga, Charles
Heron, Andrew
Seddon, John M.
Templer, Richard H.
Hankamer, Ben
Title Membrane-protein crystallization in cubo: Temperature-dependent phase behaviour of monoolein-detergent mixtures
Journal name Acta Crystallographica Section D: Biological Crystallography   Check publisher's open access policy
ISSN 0907-4449
1399-0047
Publication date 2003-02
Sub-type Article (original research)
DOI 10.1107/S0907444902020772
Open Access Status File (Publisher version)
Volume 59
Issue 2
Start page 239
End page 246
Total pages 8
Editor G. Kostorz
Place of publication Malden, MA, United States
Publisher Wiley-Blackwell
Collection year 2003
Language eng
Subject C1
270104 Membrane Biology
730299 Public health not elsewhere classified
0601 Biochemistry and Cell Biology
Formatted abstract
The lipidic cubic phase of monoolein has proved to be a matrix well suited to the production of three-dimensional crystals of membrane proteins. It consists of a single continuous bilayer, which is contorted in three-dimensional space and separates two distinct water channels. It has previously been proposed that on the addition of precipitants, membrane proteins embedded in the cubic phase migrate through the matrix to nucleation sites and that this process is dependent upon the stability of the lipidic cubic phase. Here, the effect of detergent type (C8-C12 glucosides, C8-C12 maltosides and C7 thioglucoside) and concentration (1-3x the critical micelle concentration; CMC) on cubic phase stability are reported in the form of the temperature-dependent phase behaviour (268-313 K) in 40% aqueous solution. The results are tabulated to show the best monoolein (MO)-detergent mixtures, mixing temperatures and crystallization temperatures identified. Monoolein-detergent mixtures suited for low-temperature in cubo crystallization of temperature-sensitive proteins are also reported for the first time. These mixtures can be prepared at low temperatures (mixed at ≤288 K) and remain stable at 277 K for a period of at least one month. They include MO-heptyl thioglucoside (1x and 3x CMC), MO-nonyl glucoside (3x CMC), MO-octyl maltoside (3x CMC), MO-nonyl maltoside (1x CMC) and MO-decyl maltoside (1x CMC).
Keyword Biochemical research methods
Biochemistry and molecular biology
Biophysics
Crystallography
Angstrom resolution
Bacteriorhodopsin
Mesophases
Bilayers
System
Q-Index Code C1

 
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Created: Wed, 15 Aug 2007, 02:54:02 EST