Baculovirus-driven expression and purification of glycine receptor α1 homo-oligomers

Joachim Morr; Nils Rundström; Heinrich Betz; Dieter Langosch; Bertram Schmitt

doi:10.1016/0014-5793(95)00721-K

Baculovirus-driven expression and purification of glycine receptor α1 homo-oligomers

Joachim Morr, Nils Rundström, Heinrich Betz, Dieter Langosch, Bertram Schmitt

Max-Planck-Inst fur Hirnforschung

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

13 Scopus citations

Abstract

The glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. We expressed a homo-oligomeric receptor composed of human α1 subunits in Spodoptera frugiperda cells by infection with a recombinant Autographa californica nuclear polyhedrosis virus. A substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4-to 30-fold higher than in other eukaryotic heterologous expression systems or native rat spinal cord membranes, respectively. Upon detergent solubilization, the al receptor was found to exist in a predominantly monodisperse state and could be affinity-purified to near homogeneity. This preparation is a potential starting point for future crystallisation studies.

Original language	English
Pages (from-to)	495-499
Number of pages	5
Journal	FEBS Letters
Volume	368
Issue number	3
DOIs	https://doi.org/10.1016/0014-5793(95)00721-K
State	Published - 24 Jul 1995
Externally published	Yes

Keywords

Affinity purification
Baculovirus
Glycine receptor
Heterologous expression

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10.1016/0014-5793(95)00721-K

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title = "Baculovirus-driven expression and purification of glycine receptor α1 homo-oligomers",

abstract = "The glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. We expressed a homo-oligomeric receptor composed of human α1 subunits in Spodoptera frugiperda cells by infection with a recombinant Autographa californica nuclear polyhedrosis virus. A substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4-to 30-fold higher than in other eukaryotic heterologous expression systems or native rat spinal cord membranes, respectively. Upon detergent solubilization, the al receptor was found to exist in a predominantly monodisperse state and could be affinity-purified to near homogeneity. This preparation is a potential starting point for future crystallisation studies.",

keywords = "Affinity purification, Baculovirus, Glycine receptor, Heterologous expression",

author = "Joachim Morr and Nils Rundstr{\"o}m and Heinrich Betz and Dieter Langosch and Bertram Schmitt",

note = "Funding Information: Acknowledgements: We thank Dr. A. Kirsch for help with immunofluorescence microscopy. This work was supported by the Deutsche Forschungsgemeinschaft (Leibniz-Program and SFB 169) and the Fonds der Chemischen Industrie.",

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doi = "10.1016/0014-5793(95)00721-K",

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T1 - Baculovirus-driven expression and purification of glycine receptor α1 homo-oligomers

AU - Morr, Joachim

AU - Rundström, Nils

AU - Betz, Heinrich

AU - Langosch, Dieter

AU - Schmitt, Bertram

N1 - Funding Information: Acknowledgements: We thank Dr. A. Kirsch for help with immunofluorescence microscopy. This work was supported by the Deutsche Forschungsgemeinschaft (Leibniz-Program and SFB 169) and the Fonds der Chemischen Industrie.

PY - 1995/7/24

Y1 - 1995/7/24

N2 - The glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. We expressed a homo-oligomeric receptor composed of human α1 subunits in Spodoptera frugiperda cells by infection with a recombinant Autographa californica nuclear polyhedrosis virus. A substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4-to 30-fold higher than in other eukaryotic heterologous expression systems or native rat spinal cord membranes, respectively. Upon detergent solubilization, the al receptor was found to exist in a predominantly monodisperse state and could be affinity-purified to near homogeneity. This preparation is a potential starting point for future crystallisation studies.

AB - The glycine receptor is a ligand-gated anion channel protein of postsynaptic membranes. We expressed a homo-oligomeric receptor composed of human α1 subunits in Spodoptera frugiperda cells by infection with a recombinant Autographa californica nuclear polyhedrosis virus. A substantial fraction of the recombinant receptor was incorporated as a functional channel protein into the cell's plasma membrane at expression levels 4-to 30-fold higher than in other eukaryotic heterologous expression systems or native rat spinal cord membranes, respectively. Upon detergent solubilization, the al receptor was found to exist in a predominantly monodisperse state and could be affinity-purified to near homogeneity. This preparation is a potential starting point for future crystallisation studies.

KW - Affinity purification

KW - Baculovirus

KW - Glycine receptor

KW - Heterologous expression

UR - https://www.scopus.com/pages/publications/0029151310

U2 - 10.1016/0014-5793(95)00721-K

DO - 10.1016/0014-5793(95)00721-K

M3 - Article

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AN - SCOPUS:0029151310

SN - 0014-5793

VL - 368

SP - 495

EP - 499

JO - FEBS Letters

JF - FEBS Letters

IS - 3

ER -

Baculovirus-driven expression and purification of glycine receptor α1 homo-oligomers

Abstract

Keywords

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