Journal
JOURNAL OF BIOMOLECULAR NMR
Volume 60, Issue 2-3, Pages 85-90Publisher
SPRINGER
DOI: 10.1007/s10858-014-9859-6
Keywords
Proton-detection; Sparse sampling; MAS solid-state NMR; (Semi-) automatic backbone assignment; Type III secretion needles
Categories
Funding
- Max Planck Society
- Leibniz-Institut fur Molekulare Pharmakologie
- DFG [CRC803]
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We introduce an efficient approach for sequential protein backbone assignment based on two complementary proton-detected 4D solid-state NMR experiments that correlate /N-i with CA(i)/COi or CA(i-1)/COi-1. The resulting 4D spectra exhibit excellent sensitivity and resolution and are amenable to (semi-)automatic assignment approaches. This strategy allows to obtain sequential connections with high confidence as problems related to peak overlap and multiple assignment possibilities are avoided. Non-uniform sampling schemes were implemented to allow for the acquisition of 4D spectra within a few days. Rather moderate hardware requirements enable the successful demonstration of the method on deuterated type III secretion needles using a 600 MHz spectrometer at a spinning rate of 25 kHz.
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