The membrane transporter lactose permease increases lipid bilayer bending rigidity

被引:8
作者
Mora, Nestor Lopez [1 ]
Findlay, Heather E. [1 ]
Brooks, Nicholas J. [2 ]
Purushothaman, Sowmya [2 ,3 ]
Ces, Oscar [2 ]
Booth, Paula J. [1 ]
机构
[1] Kings Coll London, Dept Chem, London, England
[2] Imperial Coll London, Dept Chem, London, England
[3] Meat, El Segundo, CA USA
基金
英国工程与自然科学研究理事会; 英国惠康基金; 欧洲研究理事会;
关键词
VESICLE FORMATION; GIANT VESICLES; ELASTICITY; PROTEINS; FLUCTUATIONS; ASYMMETRY; STABILITY; LIPOSOMES; STIFFNESS; REMOVAL;
D O I
10.1016/j.bpj.2021.06.038
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Cellular life relies on membranes, which provide a resilient and adaptive cell boundary. Many essential processes depend upon the ease with which the membrane is able to deform and bend, features that can be characterized by the bending rigidity. Quantitative investigations of such mechanical properties of biological membranes have primarily been undertaken in solely lipid bilayers and frequently in the absence of buffers. In contrast, much less is known about the influence of integral membrane proteins on bending rigidity under physiological conditions. We focus on an exemplar member of the ubiquitous major facilitator superfamily of transporters and assess the influence of lactose permease on the bending rigidity of lipid bilayers. Fluctuation analysis of giant unilamellar vesicles (GUVs) is a useful means to measure bending rigidity. We find that using a hydrogel substrate produces GUVs that are well suited to fluctuation analysis. Moreover, the hydrogel method is amenable to both physiological salt concentrations and anionic lipids, which are important to mimic key aspects of the native lactose permease membrane. Varying the fraction of the anionic lipid in the lipid mixture DOPC/DOPE/DOPG allows us to assess the dependence of membrane bending rigidity on the topology and concentration of an integral membrane protein in the lipid bilayer of GUVs. The bending rigidity gradually increases with the incorporation of lactose permease, but there is no further increase with greater amounts of the protein in the membrane.
引用
收藏
页码:3787 / 3794
页数:8
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