scholarly journals Acoustic Methods to Monitor Protein Crystallization and to Detect Protein Crystals in Suspensions of Agarose and Lipidic Cubic Phase

2016 ◽  
Vol 21 (1) ◽  
pp. 107-114 ◽  
Author(s):  
Daniel L. Ericson ◽  
Xingyu Yin ◽  
Alexander Scalia ◽  
Yasmin N. Samara ◽  
Richard Stearns ◽  
...  
Keyword(s):  
Protein Crystallization ◽  
Protein Crystals ◽  
Cubic Phase ◽  
Acoustic Methods ◽  
Lipidic Cubic Phase

scholarly journals Microscope detection options for colorless protein crystals grown in lipidic cubic phases

2003 ◽  
Vol 36 (5) ◽  
pp. 1295-1296 ◽  
Author(s):  
Peter Nollert
Keyword(s):  
Membrane Proteins ◽  
Crystal Nucleation ◽  
Protein Crystal ◽  
Protein Crystals ◽  
Cubic Phase ◽  
Polarization Microscopy ◽  
Cubic Phases ◽  
Lipidic Cubic Phase ◽  
Surface Distortions ◽  
Microscopic Techniques

The use of lipidic cubic phases as crystal nucleation and growth matrices is becoming popular and has yielded crystals of soluble and membrane proteins. So far, all of the membrane proteins crystallized by this method have been colored. This feature has facilitated the detection of the often encountered microcrystals in initial screening rounds. Indeed, small colorless protein crystals have poor optical contrast as a result of the small differences in refractive index of the protein crystal and the surrounding lipidic cubic phase. While a perfect preparation of a lipidic cubic phase is transparent and optically isotropic, in a crystallization setup it frequently disguises crystals due to cracks, inclusions, surface distortions and phase boundaries. Here, several specialized microscopic techniques and illumination conditions are compared and it is found that sufficient contrast is generated by cross polarization microscopy and by Hoffman modulation contrast microscopy for the detection of colorless protein crystals.

scholarly journals A fast, simple and robust protocol for growing crystals in the lipidic cubic phase

2012 ◽  
Vol 45 (6) ◽  
pp. 1330-1333 ◽  
Author(s):  
Margaret Aherne ◽  
Joseph A. Lyons ◽  
Martin Caffrey
Keyword(s):  
Quality Control ◽  
Data Collection ◽  
Protein Crystallization ◽  
Cubic Phase ◽  
Sandwich Plates ◽  
Protein Targets ◽  
Test Protein ◽  
Membrane Protein Crystallization ◽  
Lipidic Cubic Phase ◽  
Robust Protocol

A simple and inexpensive protocol for producing crystals in the sticky and viscous mesophase used for membrane protein crystallization by thein mesomethod is described. It provides crystals that appear within 15–30 min of setup at 293 K. The protocol gives the experimenter a convenient way of gaining familiarity and a level of comfort with the lipidic cubic mesophase, which can be daunting as a material when first encountered. Having used the protocol to produce crystals of the test protein, lysozyme, the experimenter can proceed with confidence to apply the method to more valuable membrane (and soluble) protein targets. The glass sandwich plates prepared using this robust protocol can further be used to practice harvesting and snap-cooling ofin meso-grown crystals, to explore diffraction data collection with mesophase-embedded crystals, and for an assortment of quality control and calibration applications when used in combination with a crystallization robot.

From test tube to plate: a simple procedure for the rapid preparation of microcrystallization experiments using the cubic phase method

2002 ◽  
Vol 35 (5) ◽  
pp. 637-640 ◽  
Author(s):  
Peter Nollert
Keyword(s):  
Liquid Crystalline ◽  
Simple Procedure ◽  
Protein Crystallization ◽  
Test Tube ◽  
Phase Method ◽  
Cubic Phase ◽  
Rapid Preparation ◽  
Lipidic Cubic Phase ◽  
Crystallization Conditions ◽  
Set Up

Crystals of transmembrane proteins for X-ray diffraction experiments may be grown either by employing mixed protein–detergent complexes, or in a matrix of liquid-crystalline membraneous material forming a lipidic cubic phase (in cubo). Widespread use of thein cubomethod has been severely hampered by its tediousness and the large amounts of protein required. Here a simple procedure is presented that by virtue of its simplicity and small setup size substantially reduces the preparation time as well as the amount of protein. Crystallization trials are set up in conventional multi-well plates using a semi-automatic dispenser-driven microsyringe. The microprocedure is amenable to full automation and further miniaturization. Its feasibility is demonstrated by screening for new crystallization conditions for bacteriorhodopsin using volumes ofca200 nl of lipidic cubic phase. New crystallization conditions were identified that avoid the necessity of weighing solid precipitation agents.

An efficient nanolitre-volume multi-channel device for highly viscous materials used in membrane protein crystallization

2013 ◽  
Vol 46 (3) ◽  
pp. 829-831
Author(s):  
Jinghui Luo ◽  
Raphaël Zwier ◽  
Jan Pieter Abrahams
Keyword(s):  
Lower Bound ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Crystal Structures ◽  
Protein Crystallization ◽  
Cubic Phase ◽  
Operation Speed ◽  
Membrane Protein Crystallization ◽  
Lipidic Cubic Phase ◽  
Materials Used

The crystal structures of various important membrane proteins could not have been solved without lipidic cubic phase (LCP) crystallization, and yet, compared to traditionalin surfocrystallization, LCP crystallization is not widely used because its extreme viscosity makes the cubic phase difficult to handle. Robots that can dispense LCPs are very specialized and therefore very expensive. Here, an accurate multi-channel device is described. It dispenses LCPs onto glass plates down to volumes of 20 nl accuracy and has an accuracy of 10% when dispensing 200 nl – the lower bound of LCP volumes dispensed for crystallization trials. Because of its multi-channel tips, operation speed goes up by a factor of four compared to simpler devices. It can be operated by hand, but its design also allows it to be built into a basic dispensing robot. Thus, the device lowers the threshold for LCP crystallization of membrane proteins/peptides.

scholarly journals Exploring the structural relationship between encapsulated antimicrobial peptides and the bilayer membrane mimetic lipidic cubic phase: studies with gramicidin A′

RSC Advances ◽  
2016 ◽  
Vol 6 (73) ◽  
pp. 68685-68694 ◽  
Author(s):  
Thomas G. Meikle ◽  
Charlotte E. Conn ◽  
Frances Separovic ◽  
Calum J. Drummond
Keyword(s):  
Antimicrobial Peptides ◽  
Low Cost ◽  
Bilayer Membrane ◽  
Structural Relationship ◽  
Gramicidin A ◽  
Cubic Phase ◽  
Membrane Peptides ◽  
Membrane Mimetic ◽  
Lipidic Cubic Phase ◽  
Bicontinuous Cubic

Lipid based bicontinuous cubic mesophases provide a low-cost, robust membrane mimetic nanomaterial which allows for the incorporation of membrane peptides and proteins.

scholarly journals The Lipidic Cubic Phase as a Membrane Mimetic

2011 ◽  
Vol 100 (8) ◽  
pp. 2075 ◽  
Author(s):  
Nicole Höfer ◽  
David Aragão ◽  
Martin Caffrey
Keyword(s):  
Cubic Phase ◽  
Membrane Mimetic ◽  
Lipidic Cubic Phase
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