A new approach for obtaining sequential assignment of large proteins.
A novel NMR experiment for obtaining sequential assignment of large proteins and protein complexes is described. The proposed method takes full advantage of transverse relaxation optimized spectroscopy (TROSY) and utilizes spin-state-selection to distinguish between intraresidual and sequential connectivities in the HNCA-TROSY-type correlation experiment. Thus, the intra- and interresidual cross peaks can be identified without relaying magnetization via carbonyl carbon, which relaxes very rapidly at the high magnetic fields where TROSY is most efficient. In addition, the presented method enables measurement of several scalar and residual dipolar couplings, which can potentially be used for structure determination of large proteins.
Permi
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Institute of Biotechnology, NMR Laboratory, University of Helsinki, Finland. Perttu.Permi@helsinki.fi
Annila
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D000192:Actinomycetales / Q000201:enzymology; D002247:Carbon Isotopes; D002480:Cellulase / Q000737:chemistry; D003903:Deuterium; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D003903:Deuterium; C402806:endocellulase E2; D002480:Cellulase