Quantitative Single-Molecule Analysis of Ryanodine Receptor 2 Subunit Assembly in Cardiac and Neuronal Tissues

Qianye Yin; Surya P. Aryal; Yongwook Song; Xu Fu; Christopher I. Richards

doi:10.1021/acs.analchem.4c03314

Quantitative Single-Molecule Analysis of Ryanodine Receptor 2 Subunit Assembly in Cardiac and Neuronal Tissues

Qianye Yin
, Surya P. Aryal
, Yongwook Song
, Xu Fu
, Christopher I. Richards

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

We developed a method for ex vivo receptor encapsulation and single-molecule imaging techniques from neuronal and cardiac tissues, illustrating the method’s broad applicability for measuring membrane receptor assembly. Ryanodine receptor 2 (RyR2) is a tetrameric Ca²⁺ channel governing intracellular Ca²⁺ dynamics, which is critical for muscle contraction. Employing GFP-RyR2 knock-in mice, we isolated individual receptor proteins in tissue-specific nanovesicles and performed subunit counting analyses to yield quantitative assessment of stoichiometric distributions across different organs. With this method, we explored the potential heterogeneity of brain-derived RyR2, which has been reported to form heteromeric assemblies with other ryanodine receptor isoforms.

Original language	English
Pages (from-to)	16298-16306
Number of pages	9
Journal	Analytical Chemistry
Volume	96
Issue number	41
DOIs	https://doi.org/10.1021/acs.analchem.4c03314
State	Published - Oct 15 2024

Bibliographical note

Publisher Copyright:
© 2024 American Chemical Society.

Funding

The authors acknowledge the UKY Light Microscopy Core and the Bioelectronics and Nanomedicine Center for the use of their facilities. Support for this work was provided by the NIH (GM138837 and GM138882).

Funders	Funder number
National Institutes of Health (NIH)	GM138837, GM138882

ASJC Scopus subject areas

Analytical Chemistry

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abstract = "We developed a method for ex vivo receptor encapsulation and single-molecule imaging techniques from neuronal and cardiac tissues, illustrating the method{\textquoteright}s broad applicability for measuring membrane receptor assembly. Ryanodine receptor 2 (RyR2) is a tetrameric Ca2+ channel governing intracellular Ca2+ dynamics, which is critical for muscle contraction. Employing GFP-RyR2 knock-in mice, we isolated individual receptor proteins in tissue-specific nanovesicles and performed subunit counting analyses to yield quantitative assessment of stoichiometric distributions across different organs. With this method, we explored the potential heterogeneity of brain-derived RyR2, which has been reported to form heteromeric assemblies with other ryanodine receptor isoforms.",

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AU - Song, Yongwook

AU - Fu, Xu

AU - Richards, Christopher I.

PY - 2024/10/15

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AB - We developed a method for ex vivo receptor encapsulation and single-molecule imaging techniques from neuronal and cardiac tissues, illustrating the method’s broad applicability for measuring membrane receptor assembly. Ryanodine receptor 2 (RyR2) is a tetrameric Ca2+ channel governing intracellular Ca2+ dynamics, which is critical for muscle contraction. Employing GFP-RyR2 knock-in mice, we isolated individual receptor proteins in tissue-specific nanovesicles and performed subunit counting analyses to yield quantitative assessment of stoichiometric distributions across different organs. With this method, we explored the potential heterogeneity of brain-derived RyR2, which has been reported to form heteromeric assemblies with other ryanodine receptor isoforms.

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Quantitative Single-Molecule Analysis of Ryanodine Receptor 2 Subunit Assembly in Cardiac and Neuronal Tissues

Abstract

Bibliographical note

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