Ion channel engineering using protein trans-splicing

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

Standard

Ion channel engineering using protein trans-splicing. / Sarkar, Debayan; Harms, Hendrik; Galleano, Iacopo; Sheikh, Zeshan Pervez; Pless, Stephan Alexander.

Methods in Enzymology. Elsevier, 2021. p. 19-48 (Methods in Enzymology).

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

Harvard

Sarkar, D, Harms, H, Galleano, I, Sheikh, ZP & Pless, SA 2021, Ion channel engineering using protein trans-splicing. in Methods in Enzymology. Elsevier, Methods in Enzymology, pp. 19-48. https://doi.org/10.1016/bs.mie.2021.01.028

APA

Sarkar, D., Harms, H., Galleano, I., Sheikh, Z. P., & Pless, S. A. (2021). Ion channel engineering using protein trans-splicing. In Methods in Enzymology (pp. 19-48). Elsevier. Methods in Enzymology https://doi.org/10.1016/bs.mie.2021.01.028

Vancouver

Sarkar D, Harms H, Galleano I, Sheikh ZP, Pless SA. Ion channel engineering using protein trans-splicing. In Methods in Enzymology. Elsevier. 2021. p. 19-48. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2021.01.028

Author

Sarkar, Debayan ; Harms, Hendrik ; Galleano, Iacopo ; Sheikh, Zeshan Pervez ; Pless, Stephan Alexander. / Ion channel engineering using protein trans-splicing. Methods in Enzymology. Elsevier, 2021. pp. 19-48 (Methods in Enzymology).

Bibtex

@inbook{f67ef5ff86a746eca19d250cf7d6882d,
title = "Ion channel engineering using protein trans-splicing",
abstract = "Conventional site-directed mutagenesis and genetic code expansion approaches have been instrumental in providing detailed functional and pharmacological insight into membrane proteins such as ion channels. Recently, this has increasingly been complemented by semi-synthetic strategies, in which part of the protein is generated synthetically. This means a vast range of chemical modifications, including non-canonical amino acids (ncAA), backbone modifications, chemical handles, fluorescent or spectroscopic labels and any combination of these can be incorporated. Among these approaches, protein trans-splicing (PTS) is particularly promising for protein reconstitution in live cells. It relies on one or more split inteins, which can spontaneously and covalently link flanking peptide or protein sequences. Here, we describe the use of PTS and its variant tandem PTS (tPTS) in semi-synthesis of ion channels in Xenopus laevis oocytes to incorporate ncAAs, post-translational modifications or metabolically stable mimics thereof. This strategy has the potential to expand the type and number of modifications in ion channel research.",
keywords = "Chemical modification, Membrane proteins, Protein semi-synthesis",
author = "Debayan Sarkar and Hendrik Harms and Iacopo Galleano and Sheikh, {Zeshan Pervez} and Pless, {Stephan Alexander}",
note = "Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",
year = "2021",
doi = "10.1016/bs.mie.2021.01.028",
language = "English",
isbn = "978-0-12-823924-7",
series = "Methods in Enzymology",
pages = "19--48",
booktitle = "Methods in Enzymology",
publisher = "Elsevier",
address = "Netherlands",

}

RIS

TY - CHAP

T1 - Ion channel engineering using protein trans-splicing

AU - Sarkar, Debayan

AU - Harms, Hendrik

AU - Galleano, Iacopo

AU - Sheikh, Zeshan Pervez

AU - Pless, Stephan Alexander

N1 - Publisher Copyright: © 2021 Elsevier Inc.

PY - 2021

Y1 - 2021

N2 - Conventional site-directed mutagenesis and genetic code expansion approaches have been instrumental in providing detailed functional and pharmacological insight into membrane proteins such as ion channels. Recently, this has increasingly been complemented by semi-synthetic strategies, in which part of the protein is generated synthetically. This means a vast range of chemical modifications, including non-canonical amino acids (ncAA), backbone modifications, chemical handles, fluorescent or spectroscopic labels and any combination of these can be incorporated. Among these approaches, protein trans-splicing (PTS) is particularly promising for protein reconstitution in live cells. It relies on one or more split inteins, which can spontaneously and covalently link flanking peptide or protein sequences. Here, we describe the use of PTS and its variant tandem PTS (tPTS) in semi-synthesis of ion channels in Xenopus laevis oocytes to incorporate ncAAs, post-translational modifications or metabolically stable mimics thereof. This strategy has the potential to expand the type and number of modifications in ion channel research.

AB - Conventional site-directed mutagenesis and genetic code expansion approaches have been instrumental in providing detailed functional and pharmacological insight into membrane proteins such as ion channels. Recently, this has increasingly been complemented by semi-synthetic strategies, in which part of the protein is generated synthetically. This means a vast range of chemical modifications, including non-canonical amino acids (ncAA), backbone modifications, chemical handles, fluorescent or spectroscopic labels and any combination of these can be incorporated. Among these approaches, protein trans-splicing (PTS) is particularly promising for protein reconstitution in live cells. It relies on one or more split inteins, which can spontaneously and covalently link flanking peptide or protein sequences. Here, we describe the use of PTS and its variant tandem PTS (tPTS) in semi-synthesis of ion channels in Xenopus laevis oocytes to incorporate ncAAs, post-translational modifications or metabolically stable mimics thereof. This strategy has the potential to expand the type and number of modifications in ion channel research.

KW - Chemical modification

KW - Membrane proteins

KW - Protein semi-synthesis

U2 - 10.1016/bs.mie.2021.01.028

DO - 10.1016/bs.mie.2021.01.028

M3 - Book chapter

C2 - 34120713

AN - SCOPUS:85101666696

SN - 978-0-12-823924-7

T3 - Methods in Enzymology

SP - 19

EP - 48

BT - Methods in Enzymology

PB - Elsevier

ER -

ID: 273635540