Abstract
Rotational-Echo Double-Resonance (REDOR) NMR on strategically 13C and 15N labeled samples is used to study the conformation of the LGXQ (X = S, G, or N) motif in the major ampullate gland dragline silk from the spider Nephila clavipes. A method is described for calculating REDOR dephasing curves suitable for background subtractions, using probability distributions of nitrogen atoms surrounding a given carbon site, which are developed from coordinates in the Brookhaven Protein Data Bank. The validity of the method is established by comparison to dephasings observed from natural abundance 13C peaks for G and A. Straightforward fitting of universal REDOR dephasing curves to the background corrected peaks of interest provides results which are not self-consistent, and a more sophisticated analysis is developed which better accounts for 15N labels which have scrambled from the intended positions. While there is likely some heterogeneity in the structures formed by the LGXQ sequences, the data indicate that they all form compact turn-like structures.
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