A novel high-throughput screen for identifying lipids that stabilise membrane proteins in detergent based solution.
A novel high-throughput screen for identifying lipids that stabilise membrane proteins in detergent based solution.
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DOI：
10.1371/journal.pone.0254118
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Byrne B
中科院分区：
文献类型：
--
作者：
Cecchetti C;Strauss J;Stohrer C;Naylor C;Pryor E;Hobbs J;Tanley S;Goldman A;Byrne B
膜蛋白具有一系列重要的生物学功能，并且是所有处方药物的约60%的靶点。对于大多数研究，它们需要从脂质双层中提取出来，例如通过去污剂溶解，导致天然脂质的损失，这可能会干扰重要的蛋白质-脂质/双层相互作用，从而影响功能和结构的完整性。膜蛋白的再脂质化已被证明在研究具有挑战性的靶点方面非常成功，但鉴定合适的脂质可能昂贵且费力。因此，我们开发了一种筛选方法来帮助高通量鉴定有益脂质。该屏幕覆盖了较大的脂质空间，设计适用于一系列稳定性评估方法。在这里，我们证明了它作为一种工具，用于确定稳定脂质的三种膜蛋白：细菌焦磷酸酶（Tm-PPase），真菌嘌呤转运蛋白（UapA）和人类GPCR（A2AR）。A2AR通过胆固醇半琥珀酸酯稳定，胆固醇半琥珀酸酯是一种众所周知的稳定GPCR的脂质，验证了该方法。此外，我们的筛选还确定了一系列新的脂质，这些脂质稳定了我们的测试蛋白，为进一步研究提供了起点，并证明了其作为膜蛋白研究新工具的价值。这种预先分配的屏幕将来将在商业上提供给科学界，并在该领域有许多潜在的应用。
Membrane proteins have a range of crucial biological functions and are the target of about 60% of all prescribed drugs. For most studies, they need to be extracted out of the lipid-bilayer, e.g. by detergent solubilisation, leading to the loss of native lipids, which may disturb important protein-lipid/bilayer interactions and thus functional and structural integrity. Relipidation of membrane proteins has proven extremely successful for studying challenging targets, but the identification of suitable lipids can be expensive and laborious. Therefore, we developed a screen to aid the high-throughput identification of beneficial lipids. The screen covers a large lipid space and was designed to be suitable for a range of stability assessment methods. Here, we demonstrate its use as a tool for identifying stabilising lipids for three membrane proteins: a bacterial pyrophosphatase (Tm-PPase), a fungal purine transporter (UapA) and a human GPCR (A2AR). A2AR is stabilised by cholesteryl hemisuccinate, a lipid well known to stabilise GPCRs, validating the approach. Additionally, our screen also identified a range of new lipids which stabilised our test proteins, providing a starting point for further investigation and demonstrating its value as a novel tool for membrane protein research. The pre-dispensed screen will be made commercially available to the scientific community in future and has a number of potential applications in the field.
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影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
DOI：
10.1139/bcb-2014-0076
发表时间：
2014-12-01
期刊：
BIOCHEMISTRY AND CELL BIOLOGY-BIOCHIMIE ET BIOLOGIE CELLULAIRE
影响因子：
--
作者：
Baturin, Simon;Galka, Jamie J.;O'Neil, Joe D.
通讯作者：
O'Neil, Joe D.
影响因子：
14.8
作者：
Hénault CM;Govaerts C;Spurny R;Brams M;Estrada-Mondragon A;Lynch J;Bertrand D;Pardon E;Evans GL;Woods K;Elberson BW;Cuello LG;Brannigan G;Nury H;Steyaert J;Baenziger JE;Ulens C
通讯作者：
Ulens C
影响因子：
3.8
作者：
Gourdon, Pontus;Andersen, Jacob Lauwring;Morth, Jens Preben
通讯作者：
Morth, Jens Preben
影响因子：
16.6
作者：
Jungnickel KEJ;Parker JL;Newstead S
通讯作者：
Newstead S