Less is more: A simple methyl-TROSY based pulse scheme offers improved sensitivity in applications to high molecular weight complexes

[Display omitted] •Shortened HMQC pulse sequence.•Improved sensitivity for methyl-TROSY based experiments.•Applications to complexes with molecular weights in the MDa range. The HMQC pulse sequence and variants thereof have been exploited in studies of high molecular weight protein complexes, taking...

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Bibliographic Details
Published in:Journal of magnetic resonance (1997) 2023-01, Vol.346, p.107326-107326, Article 107326
Main Authors: Bolik-Coulon, Nicolas, Sever, Alexander I.M., Harkness, Robert W., Aramini, James M., Toyama, Yuki, Hansen, D. Flemming, Kay, Lewis E.
Format: Article
Language:English
Subjects:
Carbon Isotopes
Delayed decoupling
HMQC
MDa protein complexes
Molecular Weight
Nuclear Magnetic Resonance, Biomolecular
Proteins
Sensitivity gains
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Summary:[Display omitted] •Shortened HMQC pulse sequence.•Improved sensitivity for methyl-TROSY based experiments.•Applications to complexes with molecular weights in the MDa range. The HMQC pulse sequence and variants thereof have been exploited in studies of high molecular weight protein complexes, taking advantage of the fact that fast and slow relaxing magnetization components are sequestered along two distinct magnetization transfer pathways. Despite the simplicity of the HMQC scheme an even shorter version can be designed, based on elimination of the terminal refocusing period, as a further means of increasing signal. Here we present such an experiment, and show that significant sensitivity gains, in some cases by factors of two or more, are realized in studies of proteins varying in molecular masses from 100 kDa to 1 MDa.
ISSN:1090-7807
1096-0856
DOI:10.1016/j.jmr.2022.107326