A CON-based NMR assignment strategy for pro-rich intrinsically disordered proteins with low signal dispersion: the C-terminal domain of histone H1.0 as a case study

Belén Chaves-Arquero, David Pantoja-Uceda, Alicia Roque, Inmaculada Ponte, Pedro Suau, M. Angeles Jiménez

Research output: Contribution to journalArticleResearch

5 Citations (Scopus)

Abstract

© 2018, Springer Nature B.V. The C-terminal domain of histone H1.0 (C-H1.0) is involved in DNA binding and is a main determinant of the chromatin condensing properties of histone H1.0. Phosphorylation at the (S/T)-P-X-(K/R) motifs affects DNA binding and is crucial for regulation of C-H1.0 function. Since C-H1.0 is an intrinsically disordered domain, solution NMR is an excellent approach to characterize the effect of phosphorylation on the structural and dynamic properties of C-H1.0. However, its very repetitive, low-amino acid-diverse and Pro-rich sequence, together with the low signal dispersion observed at the 1H–15N HSQC spectra of both non- and tri-phosphorylated C-H1.0 preclude the use of standard 1H-detected assignment strategies. We have achieved an essentially complete assignment of the heavy backbone atoms (15N, 13C′ and 13Cα), as well as 1HN and 13Cβ nuclei, of non- and tri-phosphorylated C-H1.0 by applying a novel 13C-detected CON-based strategy. No C-H1.0 region with a clear secondary structure tendency was detected by chemical shift analyses, confirming at residue level that C-H1.0 is disordered in aqueous solution. Phosphorylation only affected the chemical shifts of phosphorylated Thr’s, and their adjacent residues. Heteronuclear {1H}–15N NOEs were also essentially equal in the non- and tri-phosphorylated states. Hence, structural tendencies and dynamic properties of C-H1.0 free in aqueous solution are unmodified by phosphorylation. We propose that the assignment strategy used for C-H1.0, which is based on the acquisition of only a few 3D spectra, is an excellent choice for short-lived intrinsically disordered proteins with repetitive sequences.
Original languageEnglish
Pages (from-to)139-148
JournalJournal of Biomolecular NMR
Volume72
DOIs
Publication statusPublished - 1 Dec 2018

Keywords

  • Histone
  • IDP
  • Intrinsically disordered domain
  • NMR assignment strategy
  • Phosphorylation

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