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

Nicolas Bolik-Coulon; Alexander I M Sever; Robert W Harkness; James M Aramini; Yuki Toyama; D Flemming Hansen; Lewis E Kay

doi:10.1016/j.jmr.2022.107326

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

J Magn Reson. 2023 Jan:346:107326. doi: 10.1016/j.jmr.2022.107326. Epub 2022 Nov 9.

Authors

Nicolas Bolik-Coulon¹, Alexander I M Sever², Robert W Harkness¹, James M Aramini³, Yuki Toyama¹, D Flemming Hansen⁴, Lewis E Kay⁵

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.
² Department of Chemistry, University of Toronto, Toronto, ON M5S 3H6, Canada.
³ Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.
⁴ Department of Structural and Molecular Biology, Division of Biosciences, University College London, London WC1E 6BT, United Kingdom. Electronic address: d.hansen@ucl.ac.uk.
⁵ 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: 36508761
DOI: 10.1016/j.jmr.2022.107326

Abstract

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.

Keywords: Delayed decoupling; HMQC; MDa protein complexes; Sensitivity gains.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carbon Isotopes
Molecular Weight
Nuclear Magnetic Resonance, Biomolecular
Proteins*

Substances

Carbon Isotopes
Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding