脂质立方相晶体-高效筛选LCP中的结晶条件

脂质立方相(LCP)结晶已经发展成为膜蛋白结晶的一种重要方法,其中脂质单油酸甘油酯Monoolein)是创建稳定LCP的首选。然而,在过去几年中,短链脂质7.7 MAG[4,7]、7.8 MAG[6-8]和7.9 MAG[5,7]变得越来越受欢迎。最近在将高黏度脂质立方相应用于连续飞秒晶体学(LCP-SFX)方面的进展[1-3],通过以下方式进一步加速了LCP方法的成功:

  • 将载有晶体的黏性LCP直接送入X射线自由电子激光(XFEL)光束中(与液体注射器相比,减少了样品消耗);
  • 利用在LCP中生长的固有小晶体;
  • 避免了从LCP中进行棘手的晶体装载。
LCP脂质 脂质结构 货号 规格
Monoolein 9.9 MAG Monoolein 9.9 MAG LCP脂质 X-LCP-101 1 g
7.7 MAG 7.7 MAG LCP脂质 X-LCP-105 100 mg
7.8 MAG 7.8 MAG LCP脂质 X-LCP-106 100 mg
7.9 MAG
(Stable at low temperatures)
7.9 MAG LCP脂质 X-LCP-107 100 mg
Monopalmitolein 9.7 MAG Monopalmitolein 9.7 MAG LCP脂质 X-LCP-102 1 g
Monovaccenin 11.7 MAG Monovaccenin 11.7 MAG X-LCP-103 100 mg
Monoeicosenoin 11.9 MAG Monoeicosenoin 11.9 MAG X-LCP-104 1 g

 

JBScreen LCP旨在高效筛选LCP中的结晶条件。上述表格中的每种LCP脂质都能创建一个稳定的脂质立方相,可用于与JBScreen LCP进行结晶设置。其96种条件是通过对192种成功通过in meso方法结晶并产生结构的整合膜蛋白进行数据挖掘得出的[7]。该筛选按沉淀剂的浓度和类型排序,并且不含乙酸镉。

 

产品名称 货号 规格
JBScreen LCP CS-340 4×24 solutions (10 ml each)
JBScreen LCP HTS CS-213L 96 solutions (1,7 ml each)

 

文献:

[1] Nogly et al. (2016) Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography. Nat. Commun. 7:12314.

[2] Zhu et al. (2016) Serial Femtosecond Crystallography of Membrane Proteins. Adv. Exp. Med. Biol. 922:151.

[3] Batyuk et al. (2016) Native phasing of x-ray free-electron laser data for a G protein-coupled receptor. Sci. Adv. 2:e1600292.

[4] Coincon et al. (2016) Crystal structures reveal the molecular basis of ion translocation in sodium/proton antiporters. Nat. Struct. Mol. Biol. 23:248.

[5] Li et al. (2015) Ternary structure reveals mechanism of a membrane diacylglycerol kinase. Nat. Commun. 6:10140.

[6] Fowler et al. (2015) Gating Topology of the Proton-Coupled Oligopeptide Symporters. Structure 23:290.

[7] Caffrey (2015) A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes. Acta Cryst F 71:3.

[8] Li et al. (2013) Crystal structure of the integral membrane diacylglycerol kinase. Nature 497:521.

[9] Misquitta et al. (2004) Rational design of lipid for membrane protein crystallization. Journal of Structural Biology 148:169.

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