Abstract:
:TROSY-based triple resonance experiments are essential for protein backbone assignment of large biomolecular systems by solution NMR spectroscopy. In a survey of the current Bruker pulse sequence library for TROSY-based experiments we found that several sequences were plagued by artifacts that affect spectral quality and hamper data analysis. Specifically, these experiments produce sidebands in the 13C(t 1) dimension with inverted phase corresponding to 1HN resonance frequencies, with approximately 5% intensity of the parent 13C crosspeaks. These artifacts originate from the modulation of the 1HN frequency onto the resonance frequency of 13Cα and/or 13Cβ and are due to 180° pulses imperfections used for 1H decoupling during the 13C(t 1) evolution period. These sidebands can become severe for CAi, CAi-1 and/or CBi, CBi-1 correlation experiments such as TROSY-HNCACB. Here, we implement three alternative decoupling strategies that suppress these artifacts and, depending on the scheme employed, boost the sensitivity up to 14% on Bruker spectrometers. A class of comparable Agilent/Varian pulse sequences that use WALTZ16 1H decoupling can also be improved by this method resulting in up to 60-80% increase in sensitivity.
journal_name
J Biomol NMRjournal_title
Journal of biomolecular NMRauthors
Xia Y,Rossi P,Tonelli M,Huang C,Kalodimos CG,Veglia Gdoi
10.1007/s10858-017-0133-6subject
Has Abstractpub_date
2017-09-01 00:00:00pages
45-52issue
1eissn
0925-2738issn
1573-5001pii
10.1007/s10858-017-0133-6journal_volume
69pub_type
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