Investigating the Lipid Selectivity of Membrane Proteins in Heterogeneous Nanodiscs.
Investigating the Lipid Selectivity of Membrane Proteins in Heterogeneous Nanodiscs.
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DOI：
10.1021/acs.analchem.2c01488
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发表时间：
2022-06-14
影响因子：
7.4
通讯作者：
Marty, Michael T.
中科院分区：
文献类型：
--
作者：
Keener, James E.;Jayasekera, Hiruni S.;Marty, Michael T.
膜蛋白的结构和功能会受到周围脂环境的显著影响，但由于膜蛋白-脂双层的瞬时和多分散性质，膜蛋白-脂相互作用的研究往往是困难的。在这里，我们使用两种自然质谱学(MS)方法来研究大肠杆菌氨转运蛋白(AmtB)和水通道蛋白Z(AqpZ)如何选择性地重塑异质脂蛋白纳米盘中的局部脂环境。首先，我们使用气相喷射从具有不同脂类组合的异质纳米盘中分离出结合了脂类的膜蛋白。其次，我们采用溶液-洗涤剂萃取作为一种正交方法，研究了天然MS纳米盘中脂类的膜蛋白重塑。我们的结果表明，Triton X-100和LDAO保留了与气相喷射一致的脂类选择性，但C8E4扭曲了一些优先的脂类相互作用。这两种方法都表明，AmtB对磷脂酰胆碱(PC)脂类有少量选择性结合部位，对磷脂酰甘油(PG)有选择性，对磷脂酰乙醇胺(PE)无选择性。相比之下，AqpZ更喜欢PC或PG而不是PE，更喜欢PC而不是PG。总体而言，这些实验提供了一幅在混合脂质双层的背景下膜蛋白如何结合不同的脂头基团的图景。
The structure and function of membrane proteins can be significantly impacted by the surrounding lipid environment, but membrane protein-lipid interactions in lipid bilayers are often difficult to study due to their transient and polydisperse nature. Here, we used two native mass spectrometry (MS) approaches to investigate how the Escherichia coli ammonium transporter (AmtB) and aquaporin Z (AqpZ) selectively remodel their local lipid environment in heterogeneous lipoprotein nanodiscs. First, we used gas-phase ejection to isolate the membrane protein with bound lipids from heterogeneous nanodiscs with different combinations of lipids. Second, we used solution-phase detergent extraction as an orthogonal approach to study membrane protein remodeling of lipids in the nanodisc with native MS. Our results showed that Triton X-100 and LDAO retain lipid selectivity that agrees with gas-phase ejection, but C8E4 distorts some preferential lipid interactions. Both approaches reveal that AmtB has a few selective binding sites for phosphatidylcholine (PC) lipids, is selective for binding phosphatidylglycerols (PG) overall, and is nonselective for phosphatidylethanolamines (PE). In contrast, AqpZ prefers either PC or PG over PE and prefers PC over PG. Overall, these experiments provide a picture of how membrane proteins bind different lipid head groups in the context of mixed lipid bilayers.
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DOI：
10.1007/s13361-018-1976-0
发表时间：
2019-01
影响因子：
3.2
作者：
Poltash ML;McCabe JW;Patrick JW;Laganowsky A;Russell DH
通讯作者：
Russell DH
影响因子：
5.5
作者：
Bagheri Y;Ali AA;You M
通讯作者：
You M
影响因子：
16.6
作者：
Cong X;Liu Y;Liu W;Liang X;Laganowsky A
通讯作者：
Laganowsky A
影响因子：
5.4
作者：
Almen, Markus Sallman;Nordstrom, Karl J. V.;Schioth, Helgi B.
通讯作者：
Schioth, Helgi B.
影响因子：
15
作者：
Keener, James E.;Zambrano, Dane Evan;Marty, Michael T.
通讯作者：
Marty, Michael T.