Selective detection of protein acetylation by NMR spectroscopy

  • J Magn Reson. 2022 Apr:337:107169. doi: 10.1016/j.jmr.2022.107169.
Kyungryun Lee  1 ,  Sho Hee Park  1 ,  Jung Ho Lee  2
Affiliations
  • 1. Department of Chemistry, Seoul National University, Seoul 08826, South Korea.
  • 2. Department of Chemistry, Seoul National University, Seoul 08826, South Korea; Advanced Institutes of Convergence Technology, Suwon, Gyeonggi-do 16229, South Korea. Electronic address: [email protected].
PMID: 35255256 DOI: 10.1016/j.jmr.2022.107169
Abstract

Selective detection of biomolecules and their modifications in cells is essential for understanding cell functions and diseases. We have developed an NMR pulse sequence, Ac-FIND (Acetylation-FIltered aNd eDited), which uses isotope editing/filtering techniques for selective detection of protein acetylation. Acetylation of the N-terminus and lysine side chains by N-succinimidyl acetate was selectively observed for intrinsically disordered α-synuclein and well-ordered ubiquitin. Furthermore, when nonacetylated 13C/15N-enriched α-synuclein was introduced into live HEK293 cells, intracellular N-terminal acetylation of α-synuclein was detected by the appearance of a single peak using Ac-FIND. This work demonstrates the utility of NMR to detect a specific protein modification both in vitro and in live cells.

Keywords
Ac-FIND; In cell NMR; Isotope editing/filtering; Protein acetylation; α-Synuclein.
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