Crosslinking glutamate receptor ion channels

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Standard

Crosslinking glutamate receptor ion channels. / Plested, Andrew J.R.; Poulsen, Mette H.

Methods in Enzymology. Elsevier, 2021. p. 161-192 (Methods in Enzymology, Vol. 652).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Harvard

Plested, AJR & Poulsen, MH 2021, Crosslinking glutamate receptor ion channels. in Methods in Enzymology. Elsevier, Methods in Enzymology, vol. 652, pp. 161-192. https://doi.org/10.1016/bs.mie.2021.03.005

APA

Plested, A. J. R., & Poulsen, M. H. (2021). Crosslinking glutamate receptor ion channels. In Methods in Enzymology (pp. 161-192). Elsevier. Methods in Enzymology Vol. 652 https://doi.org/10.1016/bs.mie.2021.03.005

Vancouver

Plested AJR, Poulsen MH. Crosslinking glutamate receptor ion channels. In Methods in Enzymology. Elsevier. 2021. p. 161-192. (Methods in Enzymology, Vol. 652). https://doi.org/10.1016/bs.mie.2021.03.005

Author

Plested, Andrew J.R. ; Poulsen, Mette H. / Crosslinking glutamate receptor ion channels. Methods in Enzymology. Elsevier, 2021. pp. 161-192 (Methods in Enzymology, Vol. 652).

Bibtex

@inbook{7576b466dc614a42b1449e5c642066a3,
title = "Crosslinking glutamate receptor ion channels",
abstract = "Combining crosslinking strategies with electrophysiology, biochemistry, and structural in silico analysis is a powerful tool to study transient movements of ion channels during gating. This chapter describes crosslinking in living cells using cysteine and photoactive unnatural amino acids (UAAs) that we have used on glutamate receptor ion channels. Here, we share the protocol for building a perfusion tool to enable rapid chemical modification of glutamate-gated AMPA receptors, optimized for their fast activation. This system can be used to perform state-dependent crosslinking in receptors modified by cysteines or UAA incorporation on the millisecond timescale. Introducing UAAs results in receptors with lower expression levels relative to the introduction of cysteine residues. Reduced expression is principally a challenge for biochemical studies, and we share here our approach to capture the light driven oligomerization of AMPA receptors containing UAA crosslinkers. Finally, we describe strategies for computational analysis to make sense of the crosslinking results in terms of structure and function.",
keywords = "Electrophysiology, Fast perfusion, Piezo, Unnatural amino acids, UV light",
author = "Plested, {Andrew J.R.} and Poulsen, {Mette H.}",
note = "Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",
year = "2021",
doi = "10.1016/bs.mie.2021.03.005",
language = "English",
series = "Methods in Enzymology",
pages = "161--192",
booktitle = "Methods in Enzymology",
publisher = "Elsevier",
address = "Netherlands",

}

RIS

TY - CHAP

T1 - Crosslinking glutamate receptor ion channels

AU - Plested, Andrew J.R.

AU - Poulsen, Mette H.

N1 - Publisher Copyright: © 2021 Elsevier Inc.

PY - 2021

Y1 - 2021

N2 - Combining crosslinking strategies with electrophysiology, biochemistry, and structural in silico analysis is a powerful tool to study transient movements of ion channels during gating. This chapter describes crosslinking in living cells using cysteine and photoactive unnatural amino acids (UAAs) that we have used on glutamate receptor ion channels. Here, we share the protocol for building a perfusion tool to enable rapid chemical modification of glutamate-gated AMPA receptors, optimized for their fast activation. This system can be used to perform state-dependent crosslinking in receptors modified by cysteines or UAA incorporation on the millisecond timescale. Introducing UAAs results in receptors with lower expression levels relative to the introduction of cysteine residues. Reduced expression is principally a challenge for biochemical studies, and we share here our approach to capture the light driven oligomerization of AMPA receptors containing UAA crosslinkers. Finally, we describe strategies for computational analysis to make sense of the crosslinking results in terms of structure and function.

AB - Combining crosslinking strategies with electrophysiology, biochemistry, and structural in silico analysis is a powerful tool to study transient movements of ion channels during gating. This chapter describes crosslinking in living cells using cysteine and photoactive unnatural amino acids (UAAs) that we have used on glutamate receptor ion channels. Here, we share the protocol for building a perfusion tool to enable rapid chemical modification of glutamate-gated AMPA receptors, optimized for their fast activation. This system can be used to perform state-dependent crosslinking in receptors modified by cysteines or UAA incorporation on the millisecond timescale. Introducing UAAs results in receptors with lower expression levels relative to the introduction of cysteine residues. Reduced expression is principally a challenge for biochemical studies, and we share here our approach to capture the light driven oligomerization of AMPA receptors containing UAA crosslinkers. Finally, we describe strategies for computational analysis to make sense of the crosslinking results in terms of structure and function.

KW - Electrophysiology

KW - Fast perfusion

KW - Piezo

KW - Unnatural amino acids

KW - UV light

U2 - 10.1016/bs.mie.2021.03.005

DO - 10.1016/bs.mie.2021.03.005

M3 - Book chapter

C2 - 34059281

AN - SCOPUS:85103727538

T3 - Methods in Enzymology

SP - 161

EP - 192

BT - Methods in Enzymology

PB - Elsevier

ER -

ID: 273635608