Purification of A Cl--channel protein of sarcoplasmic reticulum by assaying the channel activity in the planar lipid bilayer system

Toru Ide; Hiroto Sakamoto; Takuma Morita; Takahisa Taguchi; Michiki Kasai

doi:10.1016/0006-291X(91)90886-C

Purification of A Cl^--channel protein of sarcoplasmic reticulum by assaying the channel activity in the planar lipid bilayer system

Toru Ide
, Hiroto Sakamoto
, Takuma Morita
, Takahisa Taguchi
, Michiki Kasai

Research output: Contribution to journal › Article › peer-review

17 Citations (Scopus)

Abstract

A Cl^- channel protein of sarcoplasmic reticulum (SR) was purified by assaying the channel activity in a planar lipid bilayer system. The light fraction of SR vesicles was solubilized in CHAPS and fractionated by anion exchange, gel filtration, and affinity chromatography with concanavalin A. All fractions in each step were reconstituted into vesicles with asolectin by dialysis and their channel activities were assayed after the vesicles had been fused into a planar lipid bilayer. A 100-kDa protein, different from Ca²⁺-ATPase, was found to form anion channels.

Original language	English
Pages (from-to)	38-44
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	176
Issue number	1
DOIs	https://doi.org/10.1016/0006-291X(91)90886-C
Publication status	Published - Apr 15 1991
Externally published	Yes

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0006-291X(91)90886-C

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title = "Purification of A Cl--channel protein of sarcoplasmic reticulum by assaying the channel activity in the planar lipid bilayer system",

abstract = "A Cl- channel protein of sarcoplasmic reticulum (SR) was purified by assaying the channel activity in a planar lipid bilayer system. The light fraction of SR vesicles was solubilized in CHAPS and fractionated by anion exchange, gel filtration, and affinity chromatography with concanavalin A. All fractions in each step were reconstituted into vesicles with asolectin by dialysis and their channel activities were assayed after the vesicles had been fused into a planar lipid bilayer. A 100-kDa protein, different from Ca2+-ATPase, was found to form anion channels.",

author = "Toru Ide and Hiroto Sakamoto and Takuma Morita and Takahisa Taguchi and Michiki Kasai",

note = "Funding Information: The anti-Ca*{\textquoteleft}-ATPase antibody was kindly donated by Dr. T. Yamamoto, Faculty of Science, Osaka University. This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Area of “Transmembrane Control” (02223107) and “Impulse Signaling” (02241101) to M. K. from the ministry of Education, Science and Culture of Japan.",

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doi = "10.1016/0006-291X(91)90886-C",

language = "English",

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T1 - Purification of A Cl--channel protein of sarcoplasmic reticulum by assaying the channel activity in the planar lipid bilayer system

AU - Ide, Toru

AU - Sakamoto, Hiroto

AU - Morita, Takuma

AU - Taguchi, Takahisa

AU - Kasai, Michiki

N1 - Funding Information: The anti-Ca*‘-ATPase antibody was kindly donated by Dr. T. Yamamoto, Faculty of Science, Osaka University. This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Area of “Transmembrane Control” (02223107) and “Impulse Signaling” (02241101) to M. K. from the ministry of Education, Science and Culture of Japan.

PY - 1991/4/15

Y1 - 1991/4/15

N2 - A Cl- channel protein of sarcoplasmic reticulum (SR) was purified by assaying the channel activity in a planar lipid bilayer system. The light fraction of SR vesicles was solubilized in CHAPS and fractionated by anion exchange, gel filtration, and affinity chromatography with concanavalin A. All fractions in each step were reconstituted into vesicles with asolectin by dialysis and their channel activities were assayed after the vesicles had been fused into a planar lipid bilayer. A 100-kDa protein, different from Ca2+-ATPase, was found to form anion channels.

AB - A Cl- channel protein of sarcoplasmic reticulum (SR) was purified by assaying the channel activity in a planar lipid bilayer system. The light fraction of SR vesicles was solubilized in CHAPS and fractionated by anion exchange, gel filtration, and affinity chromatography with concanavalin A. All fractions in each step were reconstituted into vesicles with asolectin by dialysis and their channel activities were assayed after the vesicles had been fused into a planar lipid bilayer. A 100-kDa protein, different from Ca2+-ATPase, was found to form anion channels.

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IS - 1

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Purification of A Cl^--channel protein of sarcoplasmic reticulum by assaying the channel activity in the planar lipid bilayer system

Abstract

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