β11-12 linker isomerization governs Acid-sensing ion channel desensitization and recovery

Rook, Matthew L.; Williamson, Abby; Lueck, John D.; Musgaard, Maria; MacLean, David M.

doi:10.1101/746271

Cited by 9 publications

(27 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ultimately testing this proposal would require further use of engineered disulfide, or possibly non‐canonical amino acid based crosslinks (Rook et al . 2020), to lock the acidic pocket open (or closed) and test the necessity and sufficiency of this domain in activation.…”

Section: The Basics Of Asicsmentioning

confidence: 99%

“…2019; Rook et al . 2020). Interestingly, the desensitization process is even sensitive to changes in the area surrounding the β11‐12 linker.…”

Section: The Basics Of Asicsmentioning

confidence: 99%

“…Reducing the hydrophobicity of this pocket by mutagenesis can profoundly accelerate both desensitization and recovery (Rook et al . 2020). Before the arrival of the cASIC1 structure, it was noted that fish and frog ASIC1 desensitized faster than their rat counter parts (Coric et al .…”

Section: The Basics Of Asicsmentioning

confidence: 99%

“…L414Bpa, and found that exposing these channels to UV light in the resting state dramatically and irreversibly reduced macroscopic desensitization (Rook et al . 2020). Taken together, all these data constitute compelling evidence that β11‐12 linker flipping is needed for channel desensitization, being akin to a switch or molecular clutch which inactivates the channel.…”

Section: The Basics Of Asicsmentioning

confidence: 99%

See 3 more Smart Citations

Coupling structure with function in acid‐sensing ion channels: challenges in pursuit of proton sensors

Rook

Musgaard

MacLean

2020

The Journal of Physiology

Self Cite

View full text Add to dashboard Cite

Acid-sensing ion channels (ASICs) are a class of trimeric cation-selective ion channels activated by changes in pH within the physiological range. They are widely expressed in the central and peripheral nervous systems where they participate in a range of physiological and pathophysiological situations such as learning and memory, pain sensation, fear and anxiety, substance abuse and cell death. ASICs are localized to cell bodies and dendrites, including the postsynaptic density, and within the last 5 years several examples of proton-evoked ASIC excitatory postsynaptic currents have emerged. Thus, ASICs have become bona fide neurotransmitter-gated ion channels, activated by the smallest neurotransmitter possible: protons. Here we review how protons are thought to drive the conformational changes associated with ASIC activation and desensitization. In particular, we weigh the evidence for and against the so-called 'acidic pocket' being a vital proton sensor and discuss the emerging role of the β11-12 linker as a desensitization switch or 'molecular clutch'. We also examine how proton-induced conformational changes pose unique challenges to classical molecular dynamics simulations, as well as some possible solutions. Matthew Rook received his Bachelor of Science degree in pharmacology and toxicology from the University at Buffalo. He is currently a PhD candidate in the laboratory of Dr David MacLean at the University of Rochester Medical Centre within the Department of Pharmacology and Physiology. His primary research project is to define the conformational changes required for acid-sensing ion channel activation and desensitization using fast-perfusion electrophysiology and genetic code expansion.

show abstract

Section: The Basics Of Asicsmentioning

confidence: 99%

“…2019; Rook et al . 2020). Interestingly, the desensitization process is even sensitive to changes in the area surrounding the β11‐12 linker.…”

Section: The Basics Of Asicsmentioning

confidence: 99%

Section: The Basics Of Asicsmentioning

confidence: 99%

Section: The Basics Of Asicsmentioning

confidence: 99%

See 2 more Smart Citations

Coupling structure with function in acid‐sensing ion channels: challenges in pursuit of proton sensors

Rook

Musgaard

MacLean

2020

The Journal of Physiology

Self Cite

View full text Add to dashboard Cite

show abstract

“…employed Bpa crosslinking to trap the ASIC1 β11‐β12 linker in distinct conformations, thereby elucidating its pivotal role in channel desensitization and recovery (Rook et al . 2020).…”

Section: Genetic Modification Of Proteinsmentioning

confidence: 99%

The current chemical biology tool box for studying ion channels

Braun

Sheikh

Pless

2020

The Journal of Physiology

View full text Add to dashboard Cite

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 analogues 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 Nina Braun studied biochemistry at the University of Bayreuth and Stockholm University before discovering her interest in ion channels and non-canonical amino acids while working on her Masters thesis with Stephan A. Pless at the University of Copenhagen. She completed her PhD in the Pless lab and is currently working on high-throughput assessment of non-canonical amino acid incorporation using photocrosslinkers in acid-sensing ion channels, with the goal of studying protein-protein interactions. Zeshan P. Sheikh completed his MSc degree at the University of Copenhagen, where he discovered his passion for ion channels, mainly focusing on recombinant expression and purification. He embarked on a PhD in the Pless lab pursuing his interest in ion channel biophysics. Here, he gained particular interest in the application of chemical biological techniques for modifying ion channels. His current research priority is applying split inteins to modify acid-sensing ion channels to gain insights into their structural dynamics.

show abstract

Molecular mechanism and structural basis of small-molecule modulation of the gating of acid-sensing ion channel 1

Liu

DesJarlais

et al. 2021

Commun Biol

View full text Add to dashboard Cite

Acid-sensing ion channels (ASICs) are proton-gated cation channels critical for neuronal functions. Studies of ASIC1, a major ASIC isoform and proton sensor, have identified acidic pocket, an extracellular region enriched in acidic residues, as a key participant in channel gating. While binding to this region by the venom peptide psalmotoxin modulates channel gating, molecular and structural mechanisms of ASIC gating modulation by small molecules are poorly understood. Here, combining functional, crystallographic, computational and mutational approaches, we show that two structurally distinct small molecules potently and allosterically inhibit channel activation and desensitization by binding at the acidic pocket and stabilizing the closed state of rat/chicken ASIC1. Our work identifies a previously unidentified binding site, elucidates a molecular mechanism of small molecule modulation of ASIC gating, and demonstrates directly the structural basis of such modulation, providing mechanistic and structural insight into ASIC gating, modulation and therapeutic targeting.

show abstract

β11-12 linker isomerization governs Acid-sensing ion channel desensitization and recovery

Cited by 9 publications

References 56 publications

Coupling structure with function in acid‐sensing ion channels: challenges in pursuit of proton sensors

Coupling structure with function in acid‐sensing ion channels: challenges in pursuit of proton sensors

The current chemical biology tool box for studying ion channels

Molecular mechanism and structural basis of small-molecule modulation of the gating of acid-sensing ion channel 1

Contact Info

Product

Resources

About