Current Recordings of Ion Channel Proteins Immobilized on Resin Beads

被引:10
作者
Hirano, Minako [1 ]
Takeuchi, Yuko [1 ]
Aoki, Takaaki [1 ]
Yanagida, Toshio [1 ]
Ide, Toru [1 ]
机构
[1] Osaka Univ, Grad Sch Frontier Biosci, Network Ctr Mol & Syst Life Sci, Suita, Osaka 5650871, Japan
关键词
POTASSIUM CHANNEL; LIPID-BILAYERS; GATING RING; MTHK; SELECTIVITY; MECHANISM; KCSA;
D O I
10.1021/ac900286z
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Current ion channel current measurement techniques are cumbersome, as they require many steps and much time. This is especially true when reconstituting channels into liposomes and incorporating them into lipid bilayers. Here, we report a novel method that measures ion channel current more efficiently than current methods. We applied our method to KcsA and MthK channels by binding them to cobalt affinity gel beads with histidine tags and then forming a lipid bilayer membrane on the bead. This allowed channels to incorporate into the bilayer and channel currents to be measured quickly and easily. The efficiency was such that currents could be recorded with extremely low amounts of protein. In addition, the channel direction could be determined by the histidine tag. This method has the potential to be applied to various channel proteins and channel research in general.
引用
收藏
页码:3151 / 3154
页数:4
相关论文
共 17 条
  • [1] Voltage-dependent gating at the KcsA selectivity filter
    Cordero-Morales, JF
    Cuello, LG
    Perozo, E
    [J]. NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2006, 13 (04) : 319 - 322
  • [2] The structure of the potassium channel:: Molecular basis of K+ conduction and selectivity
    Doyle, DA
    Cabral, JM
    Pfuetzner, RA
    Kuo, AL
    Gulbis, JM
    Cohen, SL
    Chait, BT
    MacKinnon, R
    [J]. SCIENCE, 1998, 280 (5360) : 69 - 77
  • [3] HIRANO M, 2008, SURF SCI NANOTECHNOL, V6, P130
  • [4] Direct introduction of single protein channels and pores into lipid bilayers
    Holden, MA
    Bayley, H
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2005, 127 (18) : 6502 - 6503
  • [5] An artificial lipid bilayer formed on an agarose-coated glass for simultaneous electrical and optical measurement of single ion channels
    Ide, T
    Yanagida, T
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 265 (02) : 595 - 599
  • [6] Ide T, 2002, SINGLE MOL, V3, P33, DOI 10.1002/1438-5171(200204)3:1<33::AID-SIMO33>3.0.CO
  • [7] 2-U
  • [8] Lipid bilayers at the gel interface for single ion channel recordings
    Ide, Toru
    Kobayashi, Toshihide
    Hirano, Minako
    [J]. ANALYTICAL CHEMISTRY, 2008, 80 (20) : 7792 - 7795
  • [9] Crystal structure and mechanism of a calcium-gated potassium channel
    Jiang, YX
    Lee, A
    Chen, JY
    Cadene, M
    Chait, BT
    MacKinnon, R
    [J]. NATURE, 2002, 417 (6888) : 515 - 522
  • [10] PURIFICATION AND RECONSTITUTION OF THE CALCIUM RELEASE CHANNEL FROM SKELETAL-MUSCLE
    LAI, FA
    ERICKSON, HP
    ROUSSEAU, E
    LIU, QY
    MEISSNER, G
    [J]. NATURE, 1988, 331 (6154) : 315 - 319