Immobilizing BK-channels in artificial lipid bilayers using annexin V

被引：0

作者：

Takeuchi, Yuko ^{[1
]}

Aoki, Takaaki ^{[1
]}

Yanagida, Toshio ^{[1
]}

Ide, Toru ^{[1
,2
]}

机构：

[1] Osaka Univ, Grad Sch Frontier Biosci, Soft Biosyst Grp, Labs Nanobiol, 1-3 Yamadaoka, Suita, Osaka 5650871, Japan

[2] RIKEN, Mol Informat Life Sci Res Grp, Wako, Saitama 3510198, Japan

来源：

E-JOURNAL OF SURFACE SCIENCE AND NANOTECHNOLOGY | 2007年 / 5卷 / 1-5期

关键词：

ion channel; bioimaging and engineering; molecular dynamics; electrophysiology; biophysics; immobilization;

D O I：

10.1380/ejssnt.2007.1

中图分类号：

TB3 [工程材料学];

学科分类号：

0805 ; 080502 ;

摘要：

In this report, we show an improved method for the simultaneous measurement of optical and electrical properties of single-channel proteins for analysis of the gating mechanism. Large-conductance Ca2+-activated potassium (BK) channels were isolated from porcine uterine smooth muscle and labeled with Cy5 via antibody against the N-terminal. These Cy5-labeled BK channels were incorporated into lipid bilayer membranes followed by single channel current measurements. Cy5-labeled BK channels possessed Ca2+ and voltage sensitivity. The orientation of the vesicles was determined to be outside-out. Charybdotoxin applied from the cis side blocked the channel current. For stable observations of ligand and channel binding, BK channel immobilization was also examined. The lateral diffusion coefficient of BK channels decreased over 200 fold in 1 mu M annexin V while the open probability did not change. This study is a significant advancement in simultaneous measurements of ligand binding and current change at the single channel level.

引用

页码：1 / 5

页数：5