The Effect of Detergent, Temperature, and Lipid on the Oligomeric State of MscL Constructs: Insights from Mass Spectrometry.
The Effect of Detergent, Temperature, and Lipid on the Oligomeric State of MscL Constructs: Insights from Mass Spectrometry.
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DOI：
10.1016/j.chembiol.2015.04.016
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发表时间：
2015-05-21
影响因子：
--
通讯作者：
Robinson CV
中科院分区：
文献类型：
--
作者：
Reading E;Walton TA;Liko I;Marty MT;Laganowsky A;Rees DC;Robinson CV
大电导机械敏感通道(MSCL)是细菌防止细胞溶解的渗透休克的紧急释放阀。大孔尺寸对功能至关重要，需要形成具有四聚体、五聚体或六聚体的低聚物，这取决于物种和实验方法。我们应用非变性(天然)质谱仪对5种不同的MSCL同源物在50多个不同实验条件下的寡聚状态进行了测定，阐明了脂结合和亚单位化学计量学。我们发现五聚体和四聚体之间的平衡可以被洗涤剂改变，可以被结合特定的脂类破坏，也可以被温度升高(37°C)扰乱。我们还证实存在与MSCL和其他在大肠杆菌中表达的膜蛋白结合的脂多糖，这揭示了异质性的一个潜在来源。更一般地，我们强调在与结构生物学相关的各种洗涤剂-脂类环境中使用质谱学来探测膜蛋白。
The mechanosensitive channel of large conductance (MscL) acts as an emergency release valve for osmotic shock of bacteria preventing cell lysis. The large pore size, essential for function, requires the formation of oligomers with tetramers, pentamers or hexamers observed depending on the species and experimental approach. We applied non-denaturing (native) mass spectrometry to five different homologs of MscL to determine the oligomeric state under more than 50 different experimental conditions elucidating lipid binding and subunit stoichiometry. We found equilibrium between pentameric and tetrameric species that can be altered by detergent, disrupted by binding specific lipids, and perturbed by increasing temperature (37 °C). We also established the presence of lipopolysaccharide bound to MscL and other membrane proteins expressed in Escherichia coli, revealing a potential source of heterogeneity. More generally we highlight the use of mass spectrometry in probing membrane proteins under a variety of detergent-lipid environments relevant to structural biology.
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完整膜蛋白复合物的质谱法。
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
14.8
作者：
Drew, David;Newstead, Simon;Iwata, So
通讯作者：
Iwata, So
影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1073/pnas.1106261108
发表时间：
2011-11-01
影响因子：
11.1
作者：
Ebong, Ima-obong;Morgner, Nina;Robinson, Carol V.
通讯作者：
Robinson, Carol V.
影响因子：
8
作者：
Iscla, Irene;Wray, Robin;Blount, Paul
通讯作者：
Blount, Paul