A transient post-translational modification of active site cysteine alters binding properties of the parkinsonism protein DJ-1

Arman Mussakhmetov, Igor A. Shumilin, Raushan Nugmanova, Ivan G. Shabalin, Timur Baizhumanov, Daulet Toibazar, Bekbolat Khassenov, Wladek Minor, Darkhan Utepbergenov

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

Mutations in the human protein DJ-1 cause early onset of Parkinson's disease. A reactive cysteine residue (Cys 106 ) of DJ-1 is crucial for its protective function, although the underlying mechanisms are unclear. Here we show that a fraction of bacterially expressed polyhistidine-tagged human DJ-1 could not be eluted from a Ni-nitrilotriacetate (Ni-NTA) column with 150 mM imidazole. This unusually tight binding was accompanied by the appearance of blue violet color on the Ni-NTA column. We demonstrate by X-ray crystallography that Cys 106 is carboxymethylated in a fraction of DJ-1 tightly bound to Ni-NTA and that the replacement of Cys 106 by serine abrogates the tight binding and the appearance of blue violet color. However, carboxymethylation of purified DJ-1 is insufficient to confer the tight binding to Ni-NTA. Moreover, when eluted protein was re-applied to the Ni-NTA column, no tight binding was observed, indicating that the formation of high affinity complex with Ni-NTA depends on a transient modification of Cys 106 that transforms into a Cys 106 -carboxymethyl adduct upon elution from Ni-NTA. We conclude that an unknown metabolite reacts with Cys 106 of DJ-1 to result in a transient post-translational modification. This modification is distinct from simple oxidation to sulfinic or sulfenic acids and confers altered binding properties to DJ-1 suggesting that it could serve as a signal for sensing oxidant stress.

Original languageEnglish
Pages (from-to)328-333
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume504
Issue number1
DOIs
Publication statusPublished - Sep 26 2018

Keywords

  • DJ-1
  • Oxidative stress
  • PARK7
  • Parkinson's disease
  • S-carboxymethylcysteine

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'A transient post-translational modification of active site cysteine alters binding properties of the parkinsonism protein DJ-1'. Together they form a unique fingerprint.

Cite this