Hydrazide Mimics for Protein Lysine Acylation to Assess Nucleosome Dynamics and Deubiquitinase Action

Shridhar Bhat, You Sang Hwang, Matthew D. Gibson, Michael Morgan, Sean Dixon Taverna, Yingming Zhao, Cynthia Wolberger, Michael G. Poirier, Philip A. Cole

Research output: Contribution to journalArticle

Abstract

A range of acyl-lysine (acyl-Lys) modifications on histones and other proteins have been mapped over the past decade but for most, their functional and structural significance remains poorly characterized. One limitation in the study of acyl-Lys containing proteins is the challenge of producing them or their mimics in site-specifically modified forms. We describe a cysteine alkylation-based method to install hydrazide mimics of acyl-Lys post-translational modifications (PTMs) on proteins. We have applied this method to install mimics of acetyl-Lys, 2-hydroxyisobutyryl-Lys, and ubiquityl-Lys that could be recognized selectively by relevant acyl-Lys modification antibodies. The acyl-Lys modified histone H3 proteins were reconstituted into nucleosomes to study nucleosome dynamics and stability as a function of modification type and site. We also installed a ubiquityl-Lys mimic in histone H2B and generated a diubiquitin analog, both of which could be cleaved by deubiquitinating enzymes. Nucleosomes containing the H2B ubiquityl-Lys mimic were used to study the SAGA deubiquitinating module's molecular recognition. These results suggest that acyl-Lys mimics offer a relatively simple and promising strategy to study the role of acyl-Lys modifications in the function, structure, and regulation of proteins and protein complexes.

Original languageEnglish (US)
Pages (from-to)9478-9485
Number of pages8
JournalJournal of the American Chemical Society
Volume140
Issue number30
DOIs
StatePublished - Aug 1 2018

Fingerprint

Acylation
Nucleosomes
Lysine
Proteins
Histones
Histone Code
Alkylation
Post Translational Protein Processing
Molecular recognition
Cysteine
Antibodies
Deubiquitinating Enzymes
Enzymes

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Hydrazide Mimics for Protein Lysine Acylation to Assess Nucleosome Dynamics and Deubiquitinase Action. / Bhat, Shridhar; Hwang, You Sang; Gibson, Matthew D.; Morgan, Michael; Taverna, Sean Dixon; Zhao, Yingming; Wolberger, Cynthia; Poirier, Michael G.; Cole, Philip A.

In: Journal of the American Chemical Society, Vol. 140, No. 30, 01.08.2018, p. 9478-9485.

Research output: Contribution to journalArticle

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