کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1993707 1064700 2011 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Characterization of lipid matrices for membrane protein crystallization by high-throughput small angle X-ray scattering
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Characterization of lipid matrices for membrane protein crystallization by high-throughput small angle X-ray scattering
چکیده انگلیسی

The lipidic cubic phase (LCP) has repeatedly proven to serve as a successful membrane–mimetic matrix for a variety of difficult-to-crystallize membrane proteins. While monoolein has been the predominant lipid of choice, there is a growing need for the characterization and use of other LCP host lipids, allowing exploration of a range of structural parameters such as bilayer thickness and curvature for optimal insertion, stability and crystallogenesis of membrane proteins. Here, we describe the development of a high-throughput (HT) pipeline to employ small angle X-ray scattering (SAXS) – the most direct technique to identify lipid mesophases and measure their structural parameters – to interrogate rapidly a large number of lipid samples under a variety of conditions, similar to those encountered during crystallization. Leveraging the identical setup format for LCP crystallization trials, this method allows the quickly assessment of lipid matrices for their utility in membrane protein crystallization, and could inform the tailoring of lipid and precipitant conditions to overcome specific crystallization challenges. As proof of concept, we present HT LCP–SAXS analysis of lipid samples made of monoolein with and without cholesterol, and of monovaccenin, equilibrated with solutions used for crystallization trials and LCP fluorescence recovery after photobleaching (FRAP) experiments.


► High-throughput platform for measuring structure of lipid mesophases is developed.
► The platform is useful for screening novel lipids for in meso crystallization.
► The method is tested with 3 lipid systems at a variety of crystallization conditions.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 55, Issue 4, December 2011, Pages 342–349
نویسندگان
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