scholarly journals Direct localization of detergents and bacteriorhodopsin in the lipidic cubic phase by small-angle neutron scattering

IUCrJ ◽  
2021 ◽  
Vol 8 (1) ◽  
pp. 22-32
Author(s):  
Thomas Cleveland IV ◽  
Emily Blick ◽  
Susan Krueger ◽  
Anna Leung ◽  
Tamim Darwish ◽  
...  

Lipidic cubic phase (LCP) crystallization methods have been essential in obtaining crystals of certain membrane proteins, particularly G-protein-coupled receptors. LCP crystallization is generally optimized across a large number of potential variables, one of which may be the choice of the solubilizing detergent. A better fundamental understanding of the behavior of detergents in the LCP may guide and simplify the detergent selection process. This work investigates the distribution of protein and detergent in LCP using the membrane protein bacteriorhodopsin (bR), with the LCP prepared from highly deuterated monoolein to allow contrast-matched small-angle neutron scattering. Contrast-matching allows the scattering from the LCP bilayer itself to be suppressed, so that the distribution and behavior of the protein and detergent can be directly studied. The results showed that, for several common detergents, the detergent micelle dissociates and incorporates into the LCP bilayer essentially as free detergent monomers. In addition, the detergent octyl glucoside dissociates from bR, and neither the protein nor detergent forms clusters in the LCP. The lack of detergent assemblies in the LCP implies that, upon incorporation, micelle sizes and protein/detergent interactions become less important than they would be in solution crystallization. Crystallization screening confirmed this idea, with crystals obtained from bR in the presence of most detergents tested. Thus, in LCP crystallization, detergents can be selected primarily on the basis of protein stabilization in solution, with crystallization suitability a lesser consideration.

1996 ◽  
Vol 92 (14) ◽  
pp. 2607-2610 ◽  
Author(s):  
Peter J. Hall ◽  
Wilson Ruiz Machado ◽  
David Gascon Galan ◽  
Elizabeth Barrientos Barria ◽  
David C. Sherrington

Carbon ◽  
2012 ◽  
Vol 50 (14) ◽  
pp. 5062-5075 ◽  
Author(s):  
Z. Mileeva ◽  
D.K. Ross ◽  
D. Wilkinson ◽  
S.M. King ◽  
T.A. Ryan ◽  
...  

2018 ◽  
Vol 188 ◽  
pp. 156-164 ◽  
Author(s):  
Jitendra Bahadur ◽  
Leslie F. Ruppert ◽  
Vitaliy Pipich ◽  
Richard Sakurovs ◽  
Yuri B. Melnichenko

1988 ◽  
Vol 132 ◽  
Author(s):  
J. A. Eastman ◽  
J. E. Epperson ◽  
H. Hahn ◽  
T. E. Klippert ◽  
A. Narayanasamy ◽  
...  

ABSTRACTNanophase titanium, prepared by the gas-condensation method both as aggregated powder and in lightly compacted discs, has been studied by conventional small angle neutron scattering, and by use of contrast variation methods. The contrast has been changed (a), isotopically, by means of deuterated/protonated solvents distilled into the specimen and (b) by progressive incremental oxidation of the Ti particles using fixed doses of low-pressure oxygen. It was shown that some evolution of the small angle pattern for lightly compacted nanophase Ti occurred over a period of several months at 300 K. Contrast matching by external solvent works well and has allowed the scattering lengths of oxidized and unoxidized specimens to be followed. The results imply that the scattering from metal and oxide can be separated under suitable conditions. The partial oxidation experiments indicate that there is both a fast and slow oxidation at 300 K. Also, during slow oxidation, high scattering length density scattering centers were formed whose number increased, but whose size remained the same at about 2 nm; these centers are tentatively assumed to be TiO2.


2018 ◽  
Vol 140 (40) ◽  
pp. 12720-12723 ◽  
Author(s):  
Justin Siefker ◽  
Ralf Biehl ◽  
Margarita Kruteva ◽  
Artem Feoktystov ◽  
Marc-Olivier Coppens

1988 ◽  
Vol 21 (6) ◽  
pp. 1802-1806 ◽  
Author(s):  
Yushu Matsushita ◽  
Yasushi Nakao ◽  
Ryuichi Saguchi ◽  
Katsuaki Mori ◽  
Haruhisa Choshi ◽  
...  

Langmuir ◽  
1998 ◽  
Vol 14 (12) ◽  
pp. 3182-3186 ◽  
Author(s):  
I. W. Hamley ◽  
J. A. Pople ◽  
C. Booth ◽  
Y.-W. Yang ◽  
S. M. King

2002 ◽  
Vol 41 (25) ◽  
pp. 6566-6571 ◽  
Author(s):  
Miguel Castro Diaz ◽  
Peter J. Hall ◽  
Colin E. Snape ◽  
Steven D. Brown ◽  
Ron Hughes

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