KIT | KIT-Bibliothek | Impressum | Datenschutz

Pure shift amide detection in conventional and TROSY-type experiments of $^{13}$C,$^{15}$N-labeled proteins

Haller, Jens D. 1; Bodor, Andrea; Luy, Burkhard ORCID iD icon 1
1 Institut für Organische Chemie (IOC), Karlsruher Institut für Technologie (KIT)

Abstract:

Large coupling networks in uniformly $^{13}$C,$^{15}$N-labeled biomolecules induce broad multiplets that even in flexible proteins are frequently not recognized as such. The reason is that given multiplets typically consist of a large number of individual resonances that result in a single broad line, in which individual components are no longer resolved. We here introduce a real-time pure shift acquisition scheme for the detection of amide protons which is based on 13C-BIRDr,X. As a result the full homo- and heteronuclear coupling network can be suppressed at low power leading to real singlets at substantially improved resolution and uncompromised sensitivity. The method is tested on a small globular and an intrinsically disordered protein (IDP) where the average spectral resolution is increased by a factor of ~ 2 and higher. Equally important, the approach works without saturation of water magnetization for solvent suppression and exchanging amide protons are not affected by saturation transfer.


Verlagsausgabe §
DOI: 10.5445/IR/1000153822
Veröffentlicht am 13.12.2022
Originalveröffentlichung
DOI: 10.1007/s10858-022-00406-z
Scopus
Zitationen: 2
Web of Science
Zitationen: 1
Dimensions
Zitationen: 4
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Organische Chemie (IOC)
Publikationstyp Zeitschriftenaufsatz
Publikationsmonat/-jahr 12.2022
Sprache Englisch
Identifikator ISSN: 0925-2738, 1573-5001
KITopen-ID: 1000153822
HGF-Programm 43.35.02 (POF IV, LK 01) Functionality of Soft Matter and Biomolecular Systems
Erschienen in Journal of Biomolecular NMR
Verlag Springer
Band 76
Heft 5-6
Seiten 213–221
Vorab online veröffentlicht am 18.11.2022
Nachgewiesen in Scopus
Web of Science
Dimensions
KIT – Die Forschungsuniversität in der Helmholtz-Gemeinschaft
KITopen Landing Page