Lipidic Cubic Phase-Induced Membrane Protein Crystallization: Interplay Between Lipid Molecular Structure, Mesophase Structure and Properties, and Crystallogenesis

2017 ◽  
Vol 17 (11) ◽  
pp. 5667-5674 ◽  
Author(s):  
Alexandru Zabara ◽  
Thomas G. Meikle ◽  
Raphael Trenker ◽  
Shenggen Yao ◽  
Janet Newman ◽  
...  
Keyword(s):  
Molecular Structure ◽  
Membrane Protein ◽  
Protein Crystallization ◽  
Cubic Phase ◽  
Structure And Properties ◽  
Induced Membrane ◽  
Membrane Protein Crystallization ◽  
Lipidic Cubic Phase

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 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.

scholarly journals Be Cautious with Crystal Structures of Membrane Proteins or Complexes Prepared in Detergents

Crystals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 86 ◽  
Author(s):  
Youzhong Guo
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Crystal Structures ◽  
Structure Determination ◽  
Protein Crystallization ◽  
Energy Coupling ◽  
Careful Examination ◽  
Cubic Phase ◽  
Native Cell ◽  
Membrane Protein Crystallization

Membrane proteins are an important class of macromolecules found in all living organisms and many of them serve as important drug targets. In order to understand their biological and biochemical functions and to exploit them for structure-based drug design, high-resolution and accurate structures of membrane proteins are needed, but are still rarely available, e.g., predominantly from X-ray crystallography, and more recently from single particle cryo-EM — an increasingly powerful tool for membrane protein structure determination. However, while protein-lipid interactions play crucial roles for the structural and functional integrity of membrane proteins, for historical reasons and due to technological limitations, until recently, the primary method for membrane protein crystallization has relied on detergents. Bicelle and lipid cubic phase (LCP) methods have also been used for membrane protein crystallization, but the first step requires detergent extraction of the protein from its native cell membrane. The resulting, crystal structures have been occasionally questioned, but such concerns were generally dismissed as accidents or ignored. However, even a hint of controversy indicates that methodological drawbacks in such structural research may exist. In the absence of caution, structures determined using these methods are often assumed to be correct, which has led to surprising hypotheses for their mechanisms of action. In this communication, several examples of structural studies on membrane proteins or complexes will be discussed: Resistance-Nodulation-Division (RND) family transporters, microbial rhodopsins, Tryptophan-rich Sensory Proteins (TSPO), and Energy-Coupling Factor (ECF) type ABC transporters. These analyses should focus the attention of membrane protein structural biologists on the potential problems in structure determination relying on detergent-based methods. Furthermore, careful examination of membrane proteins in their native cell environments by biochemical and biophysical techniques is warranted, and completely detergent-free systems for membrane protein research are crucially needed.

scholarly journals Membrane Protein Crystallization In Meso: Lipid Type-Tailoring of the Cubic Phase

2002 ◽  
Vol 83 (6) ◽  
pp. 3393-3407 ◽  
Author(s):  
Vadim Cherezov ◽  
Jeffrey Clogston ◽  
Yohann Misquitta ◽  
Wissam Abdel-Gawad ◽  
Martin Caffrey
Keyword(s):  
Membrane Protein ◽  
Protein Crystallization ◽  
Cubic Phase ◽  
Membrane Protein Crystallization

Phase Behavior of a Designed Cyclopropyl Analogue of Monoolein: Implications for Low-Temperature Membrane Protein Crystallization

2014 ◽  
Vol 127 (3) ◽  
pp. 1041-1045 ◽  
Author(s):  
Livia Salvati Manni ◽  
Alexandru Zabara ◽  
Yazmin M. Osornio ◽  
Jendrik Schöppe ◽  
Alexander Batyuk ◽  
...  
Keyword(s):  
Low Temperature ◽  
Membrane Protein ◽  
Phase Behavior ◽  
Protein Crystallization ◽  
Membrane Protein Crystallization
Sign in / Sign up

Export Citation Format

Share Document