Mechanism and site of action of big dynorphin on ASIC1a

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

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Mechanism and site of action of big dynorphin on ASIC1a. / Borg, Christian B.; Braun, Nina; Heusser, Stephanie A.; Bay, Yasmin; Weis, Daniel; Galleano, Iacopo; Lund, Camilla; Tian, Weihua; Haugaard-Kedström, Linda M.; Bennett, Eric P.; Lynagh, Timothy; Strømgaard, Kristian; Andersen, Jacob; Pless, Stephan A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 117, No. 13, 2020, p. 7447-7454.

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

Harvard

Borg, CB, Braun, N, Heusser, SA, Bay, Y, Weis, D, Galleano, I, Lund, C, Tian, W, Haugaard-Kedström, LM, Bennett, EP, Lynagh, T, Strømgaard, K, Andersen, J & Pless, SA 2020, 'Mechanism and site of action of big dynorphin on ASIC1a', Proceedings of the National Academy of Sciences of the United States of America, vol. 117, no. 13, pp. 7447-7454. https://doi.org/10.1073/pnas.1919323117

APA

Borg, C. B., Braun, N., Heusser, S. A., Bay, Y., Weis, D., Galleano, I., Lund, C., Tian, W., Haugaard-Kedström, L. M., Bennett, E. P., Lynagh, T., Strømgaard, K., Andersen, J., & Pless, S. A. (2020). Mechanism and site of action of big dynorphin on ASIC1a. Proceedings of the National Academy of Sciences of the United States of America, 117(13), 7447-7454. https://doi.org/10.1073/pnas.1919323117

Vancouver

Borg CB, Braun N, Heusser SA, Bay Y, Weis D, Galleano I et al. Mechanism and site of action of big dynorphin on ASIC1a. Proceedings of the National Academy of Sciences of the United States of America. 2020;117(13):7447-7454. https://doi.org/10.1073/pnas.1919323117

Author

Borg, Christian B. ; Braun, Nina ; Heusser, Stephanie A. ; Bay, Yasmin ; Weis, Daniel ; Galleano, Iacopo ; Lund, Camilla ; Tian, Weihua ; Haugaard-Kedström, Linda M. ; Bennett, Eric P. ; Lynagh, Timothy ; Strømgaard, Kristian ; Andersen, Jacob ; Pless, Stephan A. / Mechanism and site of action of big dynorphin on ASIC1a. In: Proceedings of the National Academy of Sciences of the United States of America. 2020 ; Vol. 117, No. 13. pp. 7447-7454.

Bibtex

@article{edcbbbad225248bc8654ccb3d28423d0,
title = "Mechanism and site of action of big dynorphin on ASIC1a",
abstract = "Acid-sensing ion channels (ASICs) are proton-gated cation channels that contribute to neurotransmission, as well as initiation of pain and neuronal death following ischemic stroke. As such, there is a great interest in understanding the in vivo regulation of ASICs, especially by endogenous neuropeptides that potently modulate ASICs. The most potent endogenous ASIC modulator known to date is the opioid neuropeptide big dynorphin (BigDyn). BigDyn is up-regulated in chronic pain and increases ASIC-mediated neuronal death during acidosis. Understanding the mechanism and site of action of BigDyn on ASICs could thus enable the rational design of compounds potentially useful in the treatment of pain and ischemic stroke. To this end, we employ a combination of electrophysiology, voltage-clamp fluorometry, synthetic BigDyn analogs, and noncanonical amino acid-mediated photocrosslinking. We demonstrate that BigDyn binding results in an ASIC1a closed resting conformation that is distinct from open and desensitized states induced by protons. Using alanine-substituted BigDyn analogs, we find that the BigDyn modulation of ASIC1a is primarily mediated through electrostatic interactions of basic amino acids in the BigDyn N terminus. Furthermore, neutralizing acidic amino acids in the ASIC1a extracellular domain reduces BigDyn effects, suggesting a binding site at the acidic pocket. This is confirmed by photocrosslinking using the noncanonical amino acid azidophenylalanine. Overall, our data define the mechanism of how BigDyn modulates ASIC1a, identify the acidic pocket as the binding site for BigDyn, and thus highlight this cavity as an important site for the development of ASIC-targeting therapeutics.",
author = "Borg, {Christian B.} and Nina Braun and Heusser, {Stephanie A.} and Yasmin Bay and Daniel Weis and Iacopo Galleano and Camilla Lund and Weihua Tian and Haugaard-Kedstr{\"o}m, {Linda M.} and Bennett, {Eric P.} and Timothy Lynagh and Kristian Str{\o}mgaard and Jacob Andersen and Pless, {Stephan A.}",
year = "2020",
doi = "10.1073/pnas.1919323117",
language = "English",
volume = "117",
pages = "7447--7454",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "The National Academy of Sciences of the United States of America",
number = "13",

}

RIS

TY - JOUR

T1 - Mechanism and site of action of big dynorphin on ASIC1a

AU - Borg, Christian B.

AU - Braun, Nina

AU - Heusser, Stephanie A.

AU - Bay, Yasmin

AU - Weis, Daniel

AU - Galleano, Iacopo

AU - Lund, Camilla

AU - Tian, Weihua

AU - Haugaard-Kedström, Linda M.

AU - Bennett, Eric P.

AU - Lynagh, Timothy

AU - Strømgaard, Kristian

AU - Andersen, Jacob

AU - Pless, Stephan A.

PY - 2020

Y1 - 2020

N2 - Acid-sensing ion channels (ASICs) are proton-gated cation channels that contribute to neurotransmission, as well as initiation of pain and neuronal death following ischemic stroke. As such, there is a great interest in understanding the in vivo regulation of ASICs, especially by endogenous neuropeptides that potently modulate ASICs. The most potent endogenous ASIC modulator known to date is the opioid neuropeptide big dynorphin (BigDyn). BigDyn is up-regulated in chronic pain and increases ASIC-mediated neuronal death during acidosis. Understanding the mechanism and site of action of BigDyn on ASICs could thus enable the rational design of compounds potentially useful in the treatment of pain and ischemic stroke. To this end, we employ a combination of electrophysiology, voltage-clamp fluorometry, synthetic BigDyn analogs, and noncanonical amino acid-mediated photocrosslinking. We demonstrate that BigDyn binding results in an ASIC1a closed resting conformation that is distinct from open and desensitized states induced by protons. Using alanine-substituted BigDyn analogs, we find that the BigDyn modulation of ASIC1a is primarily mediated through electrostatic interactions of basic amino acids in the BigDyn N terminus. Furthermore, neutralizing acidic amino acids in the ASIC1a extracellular domain reduces BigDyn effects, suggesting a binding site at the acidic pocket. This is confirmed by photocrosslinking using the noncanonical amino acid azidophenylalanine. Overall, our data define the mechanism of how BigDyn modulates ASIC1a, identify the acidic pocket as the binding site for BigDyn, and thus highlight this cavity as an important site for the development of ASIC-targeting therapeutics.

AB - Acid-sensing ion channels (ASICs) are proton-gated cation channels that contribute to neurotransmission, as well as initiation of pain and neuronal death following ischemic stroke. As such, there is a great interest in understanding the in vivo regulation of ASICs, especially by endogenous neuropeptides that potently modulate ASICs. The most potent endogenous ASIC modulator known to date is the opioid neuropeptide big dynorphin (BigDyn). BigDyn is up-regulated in chronic pain and increases ASIC-mediated neuronal death during acidosis. Understanding the mechanism and site of action of BigDyn on ASICs could thus enable the rational design of compounds potentially useful in the treatment of pain and ischemic stroke. To this end, we employ a combination of electrophysiology, voltage-clamp fluorometry, synthetic BigDyn analogs, and noncanonical amino acid-mediated photocrosslinking. We demonstrate that BigDyn binding results in an ASIC1a closed resting conformation that is distinct from open and desensitized states induced by protons. Using alanine-substituted BigDyn analogs, we find that the BigDyn modulation of ASIC1a is primarily mediated through electrostatic interactions of basic amino acids in the BigDyn N terminus. Furthermore, neutralizing acidic amino acids in the ASIC1a extracellular domain reduces BigDyn effects, suggesting a binding site at the acidic pocket. This is confirmed by photocrosslinking using the noncanonical amino acid azidophenylalanine. Overall, our data define the mechanism of how BigDyn modulates ASIC1a, identify the acidic pocket as the binding site for BigDyn, and thus highlight this cavity as an important site for the development of ASIC-targeting therapeutics.

U2 - 10.1073/pnas.1919323117

DO - 10.1073/pnas.1919323117

M3 - Journal article

C2 - 32165542

VL - 117

SP - 7447

EP - 7454

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 13

ER -

ID: 237511731