Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels. / Sarkar, Debayan; Galleano, Iacopo; Heusser, Stephanie Andrea; Ou, Sofie Yuewei; Uzun, Gül Refika; Khoo, Keith K.; van der Heden van Noort, Gerbrand Jan; Harrison, Joseph Scott; Pless, Stephan Alexander.

In: Cell Chemical Biology, 2024.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Sarkar, D, Galleano, I, Heusser, SA, Ou, SY, Uzun, GR, Khoo, KK, van der Heden van Noort, GJ, Harrison, JS & Pless, SA 2024, 'Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels', Cell Chemical Biology. https://doi.org/10.1016/j.chembiol.2023.11.013

APA

Sarkar, D., Galleano, I., Heusser, S. A., Ou, S. Y., Uzun, G. R., Khoo, K. K., van der Heden van Noort, G. J., Harrison, J. S., & Pless, S. A. (Accepted/In press). Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels. Cell Chemical Biology. https://doi.org/10.1016/j.chembiol.2023.11.013

Vancouver

Sarkar D, Galleano I, Heusser SA, Ou SY, Uzun GR, Khoo KK et al. Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels. Cell Chemical Biology. 2024. https://doi.org/10.1016/j.chembiol.2023.11.013

Author

Sarkar, Debayan ; Galleano, Iacopo ; Heusser, Stephanie Andrea ; Ou, Sofie Yuewei ; Uzun, Gül Refika ; Khoo, Keith K. ; van der Heden van Noort, Gerbrand Jan ; Harrison, Joseph Scott ; Pless, Stephan Alexander. / Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels. In: Cell Chemical Biology. 2024.

Bibtex

@article{ae8e1589c1ec4b5ea44dbad9a57a460e,
title = "Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels",
abstract = "Acid-sensing ion channels (ASICs) are trimeric ion channels that open a cation-conducting pore in response to proton binding. Excessive ASIC activation during prolonged acidosis in conditions such as inflammation and ischemia is linked to pain and stroke. A conserved lysine in the extracellular domain (Lys211 in mASIC1a) is suggested to play a key role in ASIC function. However, the precise contributions are difficult to dissect with conventional mutagenesis, as replacement of Lys211 with naturally occurring amino acids invariably changes multiple physico-chemical parameters. Here, we study the contribution of Lys211 to mASIC1a function using tandem protein trans-splicing (tPTS) to incorporate non-canonical lysine analogs. We conduct optimization efforts to improve splicing and functionally interrogate semisynthetic mASIC1a. In combination with molecular modeling, we show that Lys211 charge and side-chain length are crucial to activation and desensitization, thus emphasizing that tPTS can enable atomic-scale interrogations of membrane proteins in live cells.",
keywords = "acid-sensing ion channels, desensitization, homolysine, ligand-gated ion channels, native chemical ligation, non-canonical amino acids, ornithine, protein semisynthesis, protein trans-splicing, split inteins",
author = "Debayan Sarkar and Iacopo Galleano and Heusser, {Stephanie Andrea} and Ou, {Sofie Yuewei} and Uzun, {G{\"u}l Refika} and Khoo, {Keith K.} and {van der Heden van Noort}, {Gerbrand Jan} and Harrison, {Joseph Scott} and Pless, {Stephan Alexander}",
note = "Publisher Copyright: {\textcopyright} 2023 Elsevier Ltd",
year = "2024",
doi = "10.1016/j.chembiol.2023.11.013",
language = "English",
journal = "Chemistry and Biology",
issn = "2451-9448",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Protein semisynthesis underscores the role of a conserved lysine in activation and desensitization of acid-sensing ion channels

AU - Sarkar, Debayan

AU - Galleano, Iacopo

AU - Heusser, Stephanie Andrea

AU - Ou, Sofie Yuewei

AU - Uzun, Gül Refika

AU - Khoo, Keith K.

AU - van der Heden van Noort, Gerbrand Jan

AU - Harrison, Joseph Scott

AU - Pless, Stephan Alexander

N1 - Publisher Copyright: © 2023 Elsevier Ltd

PY - 2024

Y1 - 2024

N2 - Acid-sensing ion channels (ASICs) are trimeric ion channels that open a cation-conducting pore in response to proton binding. Excessive ASIC activation during prolonged acidosis in conditions such as inflammation and ischemia is linked to pain and stroke. A conserved lysine in the extracellular domain (Lys211 in mASIC1a) is suggested to play a key role in ASIC function. However, the precise contributions are difficult to dissect with conventional mutagenesis, as replacement of Lys211 with naturally occurring amino acids invariably changes multiple physico-chemical parameters. Here, we study the contribution of Lys211 to mASIC1a function using tandem protein trans-splicing (tPTS) to incorporate non-canonical lysine analogs. We conduct optimization efforts to improve splicing and functionally interrogate semisynthetic mASIC1a. In combination with molecular modeling, we show that Lys211 charge and side-chain length are crucial to activation and desensitization, thus emphasizing that tPTS can enable atomic-scale interrogations of membrane proteins in live cells.

AB - Acid-sensing ion channels (ASICs) are trimeric ion channels that open a cation-conducting pore in response to proton binding. Excessive ASIC activation during prolonged acidosis in conditions such as inflammation and ischemia is linked to pain and stroke. A conserved lysine in the extracellular domain (Lys211 in mASIC1a) is suggested to play a key role in ASIC function. However, the precise contributions are difficult to dissect with conventional mutagenesis, as replacement of Lys211 with naturally occurring amino acids invariably changes multiple physico-chemical parameters. Here, we study the contribution of Lys211 to mASIC1a function using tandem protein trans-splicing (tPTS) to incorporate non-canonical lysine analogs. We conduct optimization efforts to improve splicing and functionally interrogate semisynthetic mASIC1a. In combination with molecular modeling, we show that Lys211 charge and side-chain length are crucial to activation and desensitization, thus emphasizing that tPTS can enable atomic-scale interrogations of membrane proteins in live cells.

KW - acid-sensing ion channels

KW - desensitization

KW - homolysine

KW - ligand-gated ion channels

KW - native chemical ligation

KW - non-canonical amino acids

KW - ornithine

KW - protein semisynthesis

KW - protein trans-splicing

KW - split inteins

U2 - 10.1016/j.chembiol.2023.11.013

DO - 10.1016/j.chembiol.2023.11.013

M3 - Journal article

C2 - 38113885

AN - SCOPUS:85186212592

JO - Chemistry and Biology

JF - Chemistry and Biology

SN - 2451-9448

ER -

ID: 384876528