Biotinylated lipid bilayer disks as model membranes for biosensor analyses

被引:21
|
作者
Lundquist, Anna [1 ]
Hansen, Soren B. [1 ]
Nordstrom, Helena [2 ]
Danielson, U. Helena [2 ]
Edwards, Katarina [1 ]
机构
[1] Uppsala Univ, BMC, Dept Phys & Analyt Chem, SE-75123 Uppsala, Sweden
[2] Uppsala Univ, BMC, Dept Biochem & Organ Chem, SE-75123 Uppsala, Sweden
基金
瑞典研究理事会;
关键词
Lipid bilayer disk; Liposome; Immobilization; Streptavidin; Biotin; Biacore; Surface plasmon resonance; SPR; Biosensor; SURFACE-PLASMON RESONANCE; AGGREGATE STRUCTURE; SPHERICAL MICELLES; LIPOSOMES; PROTEIN; IMMOBILIZATION; PARTITION; MIXTURES;
D O I
10.1016/j.ab.2010.06.030
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The aim of this study was to investigate the potential of polyethylene glycol (PEG)-stabilized lipid bilayer disks as model membranes for surface plasmon resonance (SPR)-based biosensor analyses. Nanosized bilayer disks that included 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)(2000)] (DSPE-PEG(2000)-biotin) were prepared and structurally characterized by cryo-transmission electron microscopy (cryo-TEM) imaging. The biotinylated disks were immobilized via streptavidin to three different types of sensor chips (CM3, CM4, and CM5) varying in their degree of carboxymethylation and thickness of the dextran matrix. The bilayer disks were found to interact with and bind stably to the streptavidin-coated sensor surfaces. As a first step toward the use of these bilayer disks as model membranes in SPR-based studies of membrane proteins, initial investigations were carried out with cyclooxygenases I and 2 (COX 1 and COX 2). Bilayer disks were preincubated with the respective protein and thereafter allowed to interact with the sensor surface. The signal resulting from the interaction was, in both cases, significantly enhanced as compared with the signal obtained when disks alone were injected over the surface. The results of the study suggest that bilayer disks constitute a new and promising type of model membranes for SPR-based biosensor studies. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码:153 / 159
页数:7
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