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Methyl group assignment using pseudocontact shifts with PARAssign

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journal contribution
posted on 06.09.2018, 11:10 by Mathilde Lescanne, Simon P. Skinner, Anneloes Blok, Monika Timmer, Linda Cerofolini, Marco Fragai, Claudio Luchinat, Marcellus Ubbink
A new version of the program PARAssign has been evaluated for assignment of NMR resonances of the 76 methyl groups in leucines, isoleucines and valines in a 25 kDa protein, using only the structure of the protein and pseudocontact shifts (PCS) generated with a lanthanoid tag at up to three attachment sites. The number of reliable assignments depends strongly on two factors. The principle axes of the magnetic susceptibility tensors of the paramagnetic centers should not be parallel so as to avoid correlated PCS. Second, the fraction of resonances in the spectrum of a paramagnetic sample that can be paired with the diamagnetic counterparts is critical for the assignment. With the data from two tag positions a reliable assignment could be obtained for 60% of the methyl groups and for many of the remaining resonances the number of possible assignments is limited to two or three. With a single tag, reliable assignments can be obtained for methyl groups with large PCS near the tag. It is concluded that assignment of methyl group resonances by paramagnetic tagging can be particularly useful in combination with some additional data, such as from mutagenesis or NOE-based experiments. Approaches to yield the best assignment results with PCS generating tags are discussed.

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Citation

Journal of Biomolecular NMR, 2017, 69 (4), pp. 183-195

Author affiliation

/Organisation/COLLEGE OF LIFE SCIENCES/Biological Sciences/Molecular & Cell Biology

Version

VoR (Version of Record)

Published in

Journal of Biomolecular NMR

Publisher

Springer Verlag

issn

0925-2738

eissn

1573-5001

Acceptance date

25/09/2017

Copyright date

2017

Available date

06/09/2018

Publisher version

https://link.springer.com/article/10.1007/s10858-017-0136-3

Language

en

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