biostudies-literatureHaller JDHelmholtz-GemeinschaftDeutsche ForschungsgemeinschaftNemzeti Kutatási Fejlesztési és Innovációs HivatalKarlsruher Institut für Technologie (KIT)213-221https://www.ebi.ac.uk/biostudies/studies/S-EPMC9712348biostudies-literatureUnknown76(5-6)Large coupling networks in uniformly ¹³C,¹⁵N-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-BIRD^r,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.Journal of biomolecular NMRPure shift amide detection in conventional and TROSY-type experiments of ¹³C,¹⁵N-labeled proteins.PMC9712348LU 835/13-1K137940Information 43.35.02K124900Bodor AHaller JDLuy BfalsePure shift amide detection in conventional and TROSY-type experiments of ¹³C,¹⁵N-labeled proteins.Large coupling networks in uniformly ¹³C,¹⁵N-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-BIRD^r,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.2022-01-01T00:00:00Z2022 Dec2025-04-18T16:54:46.868Z2025-04-07T04:21:38.175ZS-EPMC97123483639920710.1007/s10858-022-00406-z