Backbone assignment of perdeuterated proteins using long-range H/C-dipolar transfers
 

Backbone assignment of perdeuterated proteins using long-range H/C-dipolar transfers


Abstract For micro-crystalline proteins, solid-state nuclear magnetic resonance spectroscopy of perdeuterated samples can provide spectra of unprecedented quality. Apart from allowing to detect sparsely introduced protons and thereby increasing the effective resolution for a series of sophisticated techniques, deuteration can provide extraordinary coherence lifetimesâ??obtainable for all involved nuclei virtually without decoupling and enabling the use of scalar magnetization transfers. Unfortunately, for fibrillar or membrane-embedded proteins, significantly shorter transverse relaxation times have been encountered as compared to micro-crystalline proteins despite an identical sample preparation, calling for alternative strategies for resonance assignment. In this work we propose an approach towards sequential assignment of perdeuterated proteins based on long-range 1H/13C Cross Polarization transfers. This strategy gives rise to H/N-separated correlations involving Cα, Cβ, and CO chemical shifts of both, intra- and interresidual contacts, and thus connecting adjacent residues independent of transverse relaxation times.

• Content Type Journal Article
• Category Article
• Pages 1-8
• DOI 10.1007/s10858-011-9593-2
• Authors
  • Rasmus Linser, Analytical Centre, University of New South Wales, Sydney, NSW 2052, Australia

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738

Source: Journal of Biomolecular NMR