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聚合物支撑膜中重组蛋白的摩擦阻力和电操纵

Frictional drag and electrical manipulation of recombinant proteins in polymer-supported membranes.

作者信息

Tanaka Motomu, Hermann Joachim, Haase Ilka, Fischer Markus, Boxer Steven G

机构信息

Department of Physics, Technical University of Munich, D-85748 Garching, Germany.

出版信息

Langmuir. 2007 May 8;23(10):5638-44. doi: 10.1021/la0628219. Epub 2007 Apr 5.

Abstract

We establish a lipid monolayer supported by a polymer interface that offers advantages over conventional solid-supported membranes for determining the frictional drag at the membrane-protein interface as well as for electric field manipulation of membrane-anchored proteins. Polymer-supported monolayers with functional lipid anchors allow for the specific docking of His-tagged green fluorescent protein variants (His-EGFP and His-DsRed tetramer) onto the membrane surface at a defined surface density. In the first part, we measure the lateral diffusion coefficients of lipids and proteins and calculate the frictional drag at the protein-membrane interface. The second part deals with the electric field-induced accumulation of recombinant proteins on a patterned surface. The mean drift velocity of proteins, which can be obtained analytically from the shape of the steady-state concentration gradient, can be controlled by tuning the interplay of electrophoresis and electroosmosis. The results demonstrate the potential of such molecular constructs for the local functionalization of solid substrates with membrane-associated proteins.

摘要

我们构建了一种由聚合物界面支撑的脂质单层,与传统的固体支撑膜相比,它在确定膜 - 蛋白界面的摩擦阻力以及对膜锚定蛋白进行电场操纵方面具有优势。带有功能性脂质锚定基团的聚合物支撑单层允许以确定的表面密度将His标签的绿色荧光蛋白变体(His - EGFP和His - DsRed四聚体)特异性对接至膜表面。在第一部分中,我们测量了脂质和蛋白的横向扩散系数,并计算了蛋白 - 膜界面的摩擦阻力。第二部分涉及电场诱导重组蛋白在图案化表面上的积累。蛋白的平均漂移速度可从稳态浓度梯度的形状解析得出,通过调节电泳和电渗的相互作用可以对其进行控制。结果表明了这种分子构建体在利用膜相关蛋白对固体底物进行局部功能化方面的潜力。

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