文献类型：期刊文献

英文题名：Microfluidic Diffusion Platform for Characterizing the Sizes of Lipid Vesicles and the Thermodynamics of Protein-Lipid Interactions

作者：Gang, Hongze[1,2,3];Galvagnion, Celine[1,6];Meisl, Georg[1];Muller, Thomas[1,4];Pfammatter, Manuela[1,8];Buell, Alexander K.[1,7];Levin, Aviad[1];Dobson, Christopher M.[1];Mu, Bozhong[2,3];Knowles, Tuomas P. J.[1,5]

机构：[1]Univ Cambridge, Ctr Misfolding Dis, Dept Chem, Lensfield Rd, Cambridge CB2 1EW, England;[2]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Sch Chem & Mol Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[4]Fluid Analyt Ltd, Unit 5 Chesterton Mill,Frenchs Rd, Cambridge CB4 3NP, England;[5]Univ Cambridge, Dept Phys, Cavendish Lab, JJ Thomson Ave, Cambridge CB3 1HE, England;[6]German Ctr Neurodegenerat Dis DZNE, Sigmund Freud Str 27, D-53127 Bonn, Germany;[7]Heinrich Heine Univ, Inst Phys Biol, Univ Str 1, D-40225 Dusseldorf, Germany;[8]Univ Zurich, Inst Neuropathol, CH-8091 Zurich, Switzerland

年份：2018

卷号：90

期号：5

起止页码：3284

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20181104900855);WOS:【SCI-EXPANDED(收录号:WOS:000427095700044)】；

基金：The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP7/2007-2013) through the ERC grant PhysProt (agreement No. 337969; T.P.J.K), the Newman Foundation (T.PJ.K.), the BBSRC (T.P.J.K.), The Cambridge Centre for Misfolding Diseases (C.G., C.M.D., T.P.J.K.), the Leverhulme Trust and Magdalene College, Cambridge (A.KB.), the China Scholarship Council (H.G.), and a FEBS Long-term Fellowship (A.L.).

语种：英文

外文关键词：Microfluidics - Light scattering - Biological membranes - Cytology - Cells - Size distribution - Thermodynamics - Binary mixtures - Dissociation

摘要：Elucidation of the fundamental interactions of proteins with biological membranes under native conditions is crucial for understanding the molecular basis of their biological function and malfunction. Notably, the large surface to volume ratio of living cells provides a molecular landscape for significant interactions of cellular components with membranes, thereby potentially modulating their function. However, such interactions can be challenging to probe using conventional biophysical methods due to the heterogeneity of the species and processes involved. Here, we use direct measurements of micron scale molecular diffusivity to detect and quantify the interactions of alpha-synuclein, associated with the etiology of Parkinson's disease, with negatively charged lipid vesicles. We further demonstrate that this microfluidic approach enables the characterization of size distributions of different binary mixtures of vesicles, which are not readily accessible using conventional light scattering techniques. Finally, the size distributions of the two alpha-synuclein conformations, free alpha-synuclein and membrane-bound alpha-synuclein, were resolved under varying lipid:protein ratios, thus, allowing the determination of the dissociation constant and the binding stoichiometry associated with this protein-lipid system. The microfluidic diffusional sizing platform allows these measurements to be performed on a time scale of minutes using microlitre volumes, thus, establishing the basis for an approach for the study of molecular interactions of heterogeneous systems under native conditions.