Structure and Dynamics of Membrane Proteins and Membrane Associated Proteins with Native Bicelles from Eukaryotic Tissues
Structure and Dynamics of Membrane Proteins and Membrane Associated Proteins with Native Bicelles from Eukaryotic Tissues
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真核组织天然 Bicelles 膜蛋白和膜相关蛋白的结构和动力学
DOI：
10.1021/acs.biochem.7b00575
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发表时间：
2017
期刊：
影响因子：
2.9
通讯作者：
Lorieau, Justin L.
中科院分区：
文献类型：
--
作者：
Smrt, Sean T.;Draney, Adrian W.;Singaram, Indira;Lorieau, Justin L.
在蛋白质结构、功能和动力学的体外研究中，通常排除了在活组织中发现的复杂分子相互作用范围。体内研究阐明了这些复杂的关系，但它们通常与体外广泛和受控的生物物理实验不相容。我们提出了一种替代方法，即从真核组织中提取膜以产生天然小细胞，以捕获体内研究中丰富而复杂的分子环境，同时保留体外实验的优势。从鸡蛋或小鼠大脑组织中提取的天然小细胞含有丰富的磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE）、磷脂酰甘油（PG）、磷脂酰丝氨酸（PS）、磷脂酰肌醇（PI）、磷脂酸（PA）、溶脂、胆固醇、神经酰胺（CM）和鞘磷脂（SM）。这些小细胞还含有特定来源的脂质，如三酰基甘油酯（TAGs）和分别来自鸡蛋和脑组织的硫脂。利用流感血凝素融合肽（HAfp）和淋巴细胞特异性蛋白酪氨酸激酶（lck-cSH2）的c端Src同源结构域，我们发现在天然小细胞中重组的膜蛋白和膜相关蛋白产生高分辨率核磁共振数据，并探测天然蛋白-脂质相互作用。
In vitrostudies of protein structure, function, and dynamics typically preclude the complex range of molecular interactions found in living tissues.In vivostudies elucidate these complex relationships, yet they are typically incompatible with the extensive and controlled biophysical experiments availablein vitro. We present an alternative approach by extracting membranes from eukaryotic tissues to produce native bicelles to capture the rich and complex molecular environment ofin vivostudies while retaining the advantages ofin vitroexperiments. Native bicelles derived from chicken egg or mouse cerebrum tissues contain a rich composition of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidic acid (PA), lysolipids, cholesterol, ceramides (CM), and sphingomyelin (SM). The bicelles also contain source-specific lipids such as triacylglycerides (TAGs) and sulfatides from egg and brain tissues, respectively. With the influenza hemagglutinin fusion peptide (HAfp) and the C-terminal Src homology domain of lymphocyte-specific protein-tyrosine kinase (lck-cSH2), we show that membrane proteins and membrane associated proteins reconstituted in native bicelles produce high-resolution NMR data and probe native protein–lipid interactions.