巴姆A的POTRA结构域的小角X射线散射（SAXS）表征
Small Angle X-ray Scattering (SAXS) Characterization of the POTRA Domains of BamA.
作者信息
Doerner Pamela Arden, Sousa Marcelo Carlos
机构信息
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, CO, 80309, USA.
出版信息
Methods Mol Biol. 2015;1329:149-55. doi: 10.1007/978-1-4939-2871-2_11.
BamA is the central component of the BAM complex and contains a C-terminal β-barrel domain embedded in the outer membrane, and a soluble, periplasmic domain, made out of five polypeptide transport associated (POTRA) motifs. Structural characterization of the POTRA domains was carried out by a combination of crystallographic, NMR and solution Small Angle X-ray Scattering (SAXS) approaches. Despite its limited resolution, SAXS is an excellent complement to NMR and crystallography. It is well suited to validate high-resolution models in solution and is particularly useful to characterize flexible systems such as the POTRA domains of BamA. Here we present a protocol for sample preparation and discuss the considerations of SAXS data collection and quality control, which is applicable to most soluble proteins.
BamA是BAM复合物的核心组成部分，包含一个嵌入外膜的C末端β桶结构域和一个由五个多肽转运相关（POTRA）基序组成的可溶性周质结构域。通过晶体学、核磁共振（NMR）和溶液小角X射线散射（SAXS）方法相结合，对POTRA结构域进行了结构表征。尽管SAXS分辨率有限，但它是NMR和晶体学的出色补充。它非常适合验证溶液中的高分辨率模型，对于表征诸如BamA的POTRA结构域等柔性系统特别有用。本文介绍了一种样品制备方案，并讨论了SAXS数据收集和质量控制的注意事项，该方案适用于大多数可溶性蛋白质。
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