Mistic's membrane association and its assistance in overexpression of a human GPCR are independent processes
Mistic's membrane association and its assistance in overexpression of a human GPCR are independent processes
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Mistic 的膜关联及其对人类 GPCR 过度表达的帮助是独立的过程
作者：
Marino;Bordag;Keller
枯草杆菌枯草杆菌蛋白Mistic与细菌膜的相互作用以及它在促进其他膜蛋白过度表达方面的作用仍然存在争议。在这项研究中，我们的目的是确定Mistic的单个螺旋片段是否足以在体内和体外与膜相互作用。为此，围绕每个Mistic螺旋片段及其组合的片段被生产为GFP-融合，并研究了它们在大肠杆菌中的细胞定位。此外，还利用固相合成法合成了四个螺旋片段对应的多肽，并用圆二色谱研究了它们在各种脂类和洗涤剂中获得二级结构的能力。这两种类型的实验都表明，Mistic的第三个螺旋片段只与LDAO胶束相互作用，但不分割成脂双层。有趣的是，其他三个螺旋在体内和体外都与膜相互作用。然而，所有这些短序列都可以取代全长Mistic作为N端融合，实现人类G蛋白偶联受体ine的过度表达。虽然对蛋白质产量和质量有不同的影响，但在大肠杆菌中仍有表达。对Mistic家族的生物信息学分析将同源基因的数量从4个扩大到20个，包括芽孢杆菌属外的蛋白质。这一信息使我们在操纵子X-yugO中发现了一个高度保守的Shine-Dalgarno序列，这对钾离子通道yugO的下游翻译是重要的。
The interaction of theBacillus subtilisprotein Mistic with the bacterial membrane and its role in promoting the overexpression of other membrane proteins are still matters of debate. In this study, we aimed to determine whether individual helical fragments of Mistic are sufficient for its interaction with membranesin vivoandin vitro. To this end, fragments encompassing each of Mistic's helical segments and combinations of them were produced as GFP‐fusions, and their cellular localization was studied inEscherichia coli. Furthermore, peptides corresponding to the four helical fragments were synthesized by solid‐phase peptide synthesis, and their ability to acquire secondary structure in a variety of lipids and detergents was studied by circular dichroism spectroscopy. Both types of experiments demonstrate that the third helical fragment of Mistic interacts only with LDAO micelles but does not partition into lipid bilayers. Interestingly, the other three helices interact with membranesin vivoandin vitro. Nevertheless, all of these short sequences can replace full‐length Mistic as N‐terminal fusions to achieve overexpression of a human G‐protein‐coupled receptor inE. coli, although with different effects on quantity and quality of the protein produced. A bioinformatic analysis of the Mistic family expanded the number of homologs from 4 to 20, including proteins outside the genusBacillus. This information allowed us to discover a highly conserved Shine‐Dalgarno sequence in the operonmstX‐yugOthat is important for downstream translation of the potassium ion channelyugO.
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影响因子：
1.6
作者：
Haberstock, Stefan;Roos, Christian;Bernhard, Frank
通讯作者：
Bernhard, Frank
膜特性在 Mistic 折叠和二聚化中的作用。
DOI：
--
发表时间：
2011
期刊：
Protein engineering, design & selection : PEDS
影响因子：
--
作者：
D. Debnath;R. Basaiawmoit;K. Nielsen;D. Otzen
通讯作者：
D. Otzen
影响因子：
15
作者：
Broecker, Jana;Fiedler, Sebastian;Keller, Sandro
通讯作者：
Keller, Sandro
α螺旋膜相关蛋白Mistic的变性机制和展开状态
影响因子：
15
作者：
Jacso T;Bardiaux B;Broecker J;Fiedler S;Barwinkel T;Mainz A;Fink U;Vargas C;Oschkinat H;Keller S;Reif B
通讯作者：
Reif B
影响因子：
64.8
作者：
Kunst, F;Ogasawara, N;Danchin, A
通讯作者：
Danchin, A