The current chemical biology tool box for studying ion channels
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The current chemical biology tool box for studying ion channels. / Braun, N; Sheikh, Z P; Pless, S A.
In: The Journal of Physiology, Vol. 598, No. 20, 2020, p. 4455-4471.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - The current chemical biology tool box for studying ion channels
AU - Braun, N
AU - Sheikh, Z P
AU - Pless, S A
N1 - This article is protected by copyright. All rights reserved.
PY - 2020
Y1 - 2020
N2 - Ion channels play important roles in human physiology and their dysfunction is linked to a variety of diseases. This has sparked considerable interest in their molecular function and pharmacology and generated a need to manipulate them with great precision. The use of high-sensitivity electrophysiological methods allows for the implementation of chemical biology manipulations, as even minute protein amounts can be studied. For example, modification of solvent-accessible cysteines is a powerful tool to site-selectively modify proteins through the introduction of charged moieties or those with fluorescent properties. This has been harnessed to study ion conduction pathways and monitor conformational dynamics. In ligand-directed chemistry, a high-affinity ligand is used to modify an ion channel with a chemical probe via a reactive linker. While these approaches are typically limited to extracellular positions, genetic code expansion provides a means to introduce non-canonical amino acids in any position of the protein. This enables the insertion of subtle analogs of naturally occurring side chains or the protein backbone, as well as amino acids with fluorescent, cross-linking or photo-switchable properties. Finally, protein semi-synthesis enables the simultaneous insertion of multiple modifications, including those that would not be tolerated by the ribosomal translation machinery. Collectively, these chemical biology tools have overcome various shortcomings of conventional mutagenesis and vastly expanded the scope of possible modifications and the type of ion channels they can be applied to. Their application in both heterologous and native cell systems will no doubt play an increasingly important role in ion channel research. This article is protected by copyright. All rights reserved.
AB - Ion channels play important roles in human physiology and their dysfunction is linked to a variety of diseases. This has sparked considerable interest in their molecular function and pharmacology and generated a need to manipulate them with great precision. The use of high-sensitivity electrophysiological methods allows for the implementation of chemical biology manipulations, as even minute protein amounts can be studied. For example, modification of solvent-accessible cysteines is a powerful tool to site-selectively modify proteins through the introduction of charged moieties or those with fluorescent properties. This has been harnessed to study ion conduction pathways and monitor conformational dynamics. In ligand-directed chemistry, a high-affinity ligand is used to modify an ion channel with a chemical probe via a reactive linker. While these approaches are typically limited to extracellular positions, genetic code expansion provides a means to introduce non-canonical amino acids in any position of the protein. This enables the insertion of subtle analogs of naturally occurring side chains or the protein backbone, as well as amino acids with fluorescent, cross-linking or photo-switchable properties. Finally, protein semi-synthesis enables the simultaneous insertion of multiple modifications, including those that would not be tolerated by the ribosomal translation machinery. Collectively, these chemical biology tools have overcome various shortcomings of conventional mutagenesis and vastly expanded the scope of possible modifications and the type of ion channels they can be applied to. Their application in both heterologous and native cell systems will no doubt play an increasingly important role in ion channel research. This article is protected by copyright. All rights reserved.
U2 - 10.1113/JP276695
DO - 10.1113/JP276695
M3 - Journal article
C2 - 32715480
VL - 598
SP - 4455
EP - 4471
JO - The Journal of Physiology
JF - The Journal of Physiology
SN - 0022-3751
IS - 20
ER -
ID: 245324301