Automated Resonance Assignment of Proteins: 6 DAPSY-NMR
Journal of Biomolecular NMR(2006)
摘要
The 6-dimensional(6D) APSY-seq-HNCOCANH NMR experiment correlates two sequentially neighbor in gamidemoieties in proteins via the C′ and C α nuclei, with efficient suppression of the back transfer from C α to the originating amidemoiety. The automatic analysis of two-dimensional(2D) projections of this 6D experiment with the use of GAPRO (Hilleretal., 2005) provides a high-precision 6D peak list, which permits automated sequential assignments of proteins with the assignment software GARANT (Bartels et al., 1997). The procedure was applied to two proteins, the 63-residue 434-repressor(1-3) and the 115-residue TM1290. For both proteins, complete sequential assignments for all NMR-observable backbone resonances were obtained, and the polypeptide segments thus identified could be unambiguously located in the amino acid sequence. These results demonstrate that APSY-NMR spectroscopy in combination with a suitable assignment algorithm can provide fully automated sequence-specific backbone assignments of small proteins.
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关键词
6DAPSY-seq-HNCOCANH experiment,automatic backbone NMR assignment,GAPRO software,GARANT software,protein structure determination
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