A delayed decoupling methyl-TROSY pulse sequence for atomic resolution studies of folded proteins and RNAs in condensates

doi:10.1016/j.jmr.2024.107667

. 2024 May:362:107667.

doi: 10.1016/j.jmr.2024.107667. Epub 2024 Mar 28.

A delayed decoupling methyl-TROSY pulse sequence for atomic resolution studies of folded proteins and RNAs in condensates

Rashik Ahmed¹, Atul K Rangadurai¹, Lisa Ruetz², Martin Tollinger², Christoph Kreutz², Lewis E Kay³

Affiliations

¹ Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Chemistry, University of Toronto, Toronto, ON, M5S 3H6, Canada; Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada; Program in Molecular Medicine, Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada.
² Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
³ Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Chemistry, University of Toronto, Toronto, ON, M5S 3H6, Canada; Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada; Program in Molecular Medicine, Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada. Electronic address: kay@pound.med.utoronto.ca.

PMID: 38626504
DOI: 10.1016/j.jmr.2024.107667

A delayed decoupling methyl-TROSY pulse sequence for atomic resolution studies of folded proteins and RNAs in condensates

Rashik Ahmed et al. J Magn Reson. 2024 May.

. 2024 May:362:107667.

doi: 10.1016/j.jmr.2024.107667. Epub 2024 Mar 28.

Authors

Rashik Ahmed¹, Atul K Rangadurai¹, Lisa Ruetz², Martin Tollinger², Christoph Kreutz², Lewis E Kay³

Affiliations

¹ Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Chemistry, University of Toronto, Toronto, ON, M5S 3H6, Canada; Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada; Program in Molecular Medicine, Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada.
² Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
³ Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Chemistry, University of Toronto, Toronto, ON, M5S 3H6, Canada; Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada; Program in Molecular Medicine, Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada. Electronic address: kay@pound.med.utoronto.ca.

PMID: 38626504
DOI: 10.1016/j.jmr.2024.107667

Abstract

Solution NMR spectroscopy has tremendous potential for providing atomic resolution insights into the interactions between proteins and nucleic acids partitioned into condensed phases of phase-separated systems. However, the highly viscous nature of the condensed phase challenges applications, and in particular, the extraction of quantitative, site-specific information. Here, we present a delayed decoupling-based HMQC pulse sequence for methyl-TROSY studies of 'client' proteins and nucleic acids partitioned into 'scaffold' proteinaceous phase-separated solvents. High sensitivity and excellent quality spectra are recorded of a nascent form of superoxide dismutase and of a small RNA fragment partitioned into CAPRIN1 condensates.

Keywords: CAPRIN1; Delayed decoupling; HMQC; Phase separation; Protein and RNA ‘clients’ in condensates; Sensitivity gains.

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Conflict of interest statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Lewis E. Kay is an associate editor of this journal (JMR). If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper].

Cited by

Atomic resolution map of the solvent interactions driving SOD1 unfolding in CAPRIN1 condensates.
Ahmed R, Liang M, Hudson RP, Rangadurai AK, Huang SK, Forman-Kay JD, Kay LE. Ahmed R, et al. Proc Natl Acad Sci U S A. 2024 Aug 27;121(35):e2408554121. doi: 10.1073/pnas.2408554121. Epub 2024 Aug 22. Proc Natl Acad Sci U S A. 2024. PMID: 39172789

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A delayed decoupling methyl-TROSY pulse sequence for atomic resolution studies of folded proteins and RNAs in condensates

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A delayed decoupling methyl-TROSY pulse sequence for atomic resolution studies of folded proteins and RNAs in condensates

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