小柯机器人

美国耶鲁大学Kallol Gupta团队近期取得重要工作进展，他们最新研究提出了从完整的可定制双分子层中直接测定整体膜蛋白和脂质的低聚组织的方法。相关研究成果2023年4月27日在线发表于《自然—方法学》杂志上。

据介绍，整体膜蛋白（IMP）和脂质在膜上的分级组织对于调节无数下游信号传导至关重要。对这些过程的定量理解需要直接从完整的膜中检测IMP和脂质的寡聚组织，并确定调节它们的关键膜成分和性质。

研究人员开发了一个平台，可以直接从完整和可定制的脂质膜中对IMP-脂质复合物进行天然质谱（nMS）分析。这些双层的脂质组成和膜性质（如曲率、张力和流动性）都可以根据目标膜进行精确定制。随后对这些完整的蛋白脂质囊泡的直接nMS分析可以产生嵌入的IMP的寡聚状态，鉴定结合的脂质，并确定可以调节观察到的IMP-脂质组织的膜性质。

总之，应用这种方法，研究人员展示了脂质结合如何调节神经递质的释放，以及膜组成如何调节转运蛋白的功能性寡聚状态。

附：英文原文

Title: Direct determination of oligomeric organization of integral membrane proteins and lipids from intact customizable bilayer

Author: Panda, Aniruddha, Giska, Fabian, Duncan, Anna L., Welch, Alexander J., Brown, Caroline, McAllister, Rachel, Hariharan, Parameswaran, Goder, Jean N. D., Coleman, Jeff, Ramakrishnan, Sathish, Pincet, Frdric, Guan, Lan, Krishnakumar, Shyam, Rothman, James E., Gupta, Kallol

Issue&Volume: 2023-04-27

Abstract: Hierarchical organization of integral membrane proteins (IMP) and lipids at the membrane is essential for regulating myriad downstream signaling. A quantitative understanding of these processes requires both detections of oligomeric organization of IMPs and lipids directly from intact membranes and determination of key membrane components and properties that regulate them. Addressing this, we have developed a platform that enables native mass spectrometry (nMS) analysis of IMP–lipid complexes directly from intact and customizable lipid membranes. Both the lipid composition and membrane properties (such as curvature, tension, and fluidity) of these bilayers can be precisely customized to a target membrane. Subsequent direct nMS analysis of these intact proteolipid vesicles can yield the oligomeric states of the embedded IMPs, identify bound lipids, and determine the membrane properties that can regulate the observed IMP–lipid organization. Applying this method, we show how lipid binding regulates neurotransmitter release and how membrane composition regulates the functional oligomeric state of a transporter.

DOI: 10.1038/s41592-023-01864-5

Source: https://www.nature.com/articles/s41592-023-01864-5

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex