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Journal of Biomolecular NMR

, Volume 14, Issue 2, pp 181–184 | Cite as

Measurement of 3hJNC′ connectivities across hydrogen bonds in a 30 kDa protein

  • Yun-Xing Wang
  • Jaison Jacob
  • Florence Cordier
  • Paul Wingfield
  • Stephen J. Stahl
  • Sylvia Lee-Huang
  • Dennis Torchia
  • Stephan Grzesiek
  • Ad Bax
Article

Abstract

A method is described which permits detection of 3h JNC′ scalar couplings across hydrogen bonds in larger, perdeuterated proteins. The experiment is demonstrated for the uniformly 2 H/13C/15N-enriched 30 kDa ribosome inactivating protein MAP30. The 3h JNC′ interactions are smaller than 1 Hz, but their detection in an HNCO experiment is made possible through the use of constructive interference between the 15N chemical shift anisotropy and 1 H-15N dipole-dipole relaxation mechanisms in a manner similar to that of recently proposed TROSY schemes. Sensitivity of the HNCO experiment depends strongly on the 15 N transverse relaxation rate of the downfield 15 N multiplet component and on the amide proton T1. In perdeuterated MAP30 at 40 °C, the average TROSY T2 was 169 ms at 750 MHz 1 H frequency, and a wide range of longitudinal relaxation rates was observed for the amide protons.

hydrogen bond J coupling MAP30 perdeuteration relaxation rates TROSY 

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Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Yun-Xing Wang
    • 1
  • Jaison Jacob
    • 1
  • Florence Cordier
    • 2
  • Paul Wingfield
    • 3
  • Stephen J. Stahl
    • 3
  • Sylvia Lee-Huang
    • 4
  • Dennis Torchia
    • 1
  • Stephan Grzesiek
    • 2
  • Ad Bax
    • 5
  1. 1.Molecular Structural Biology Unit, National Institute of Dental ResearchNational Institutes of HealthBethesdaU.S.A.
  2. 2.Institute of Structural BiologyForschungszentrum JülichJülichGermany
  3. 3.Protein Expression Laboratory, NationalInstitute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of HealthBethesdaU.S.A.
  4. 4.Department of BiochemistryNew York University School of MedicineNew YorkU.S.A.
  5. 5.Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaU.S.A.

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