生物素与磷脂头部基团之间不同间隔物对抗生物素蛋白-生物素相互作用促进的含生物素化磷脂脂质体固定化和沉降的影响。

Effects of various spacers between biotin and the phospholipid headgroup on immobilization and sedimentation of biotinylated phospholipid-containing liposomes facilitated by avidin-biotin interactions.

作者信息

Sakamoto Yasuhisa, Kikuchi Koji, Umeda Kazuaki, Nakanishi Hiroyuki

机构信息

Department of Molecular Pharmacology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan.

出版信息

J Biochem. 2017 Sep 1;162(3):221-226. doi: 10.1093/jb/mvx016.

DOI:10.1093/jb/mvx016

PMID:28444248

Abstract

Immobilization and sedimentation of liposomes (lipid vesicles) are used in liposome-protein binding assays, facilitated by avidin/streptavidin/NeutrAvidin and biotinylated phospholipid-containing liposomes. Here, we examined the effects of three spacers [six-carbon (X), polyethylene glycol (PEG) 180 (molecular weight 180) and PEG2000 (molecular weight 2,000)] between biotin and the phospholipid headgroup on the immobilization and sedimentation of small unilamellar liposomes/vesicles (SUVs). PEG180 and PEG2000 showed more efficient immobilization of biotinylated SUVs on NeutrAvidin-coated plates than X, but X and PEG180 showed more efficient sedimentation of biotinylated SUVs upon NeutrAvidin addition than PEG2000. Thus, the most appropriate spacers differed between immobilization and sedimentation. A spacer for biotinylated SUVs must be selected according to the particular liposome-protein binding assays examined.

摘要

脂质体（脂质小泡）的固定化和沉降用于脂质体-蛋白质结合测定，这借助于抗生物素蛋白/链霉抗生物素蛋白/中性抗生物素蛋白以及含生物素化磷脂的脂质体得以实现。在此，我们研究了生物素与磷脂头部基团之间的三种间隔臂[六碳（X）、聚乙二醇（PEG）180（分子量180）和PEG2000（分子量2000）]对小单层脂质体/小泡（SUV）的固定化和沉降的影响。PEG180和PEG2000在将生物素化SUV固定于中性抗生物素蛋白包被的平板上比X表现出更高的效率，但在加入中性抗生物素蛋白后，X和PEG180在使生物素化SUV沉降方面比PEG2000表现出更高的效率。因此，固定化和沉降时最合适的间隔臂有所不同。必须根据所检测的特定脂质体-蛋白质结合测定来选择用于生物素化SUV的间隔臂。