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Biochem Biophys Res Commun. 2018 Sep 26;504(1):328-333. doi: 10.1016/j.bbrc.2018.08.190. Epub 2018 Sep 4.

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

Author information

1
National Center for Biotechnology, Astana, 010000, Kazakhstan.
2
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, 22908, USA.
3
National Center for Biotechnology, Astana, 010000, Kazakhstan; Department of Chemistry, School of Science and Technology, Nazarbayev University, Astana, 010000, Kazakhstan.
4
Department of Chemistry, School of Science and Technology, Nazarbayev University, Astana, 010000, Kazakhstan.
5
Department of Biology, School of Science and Technology, Nazarbayev University, Astana, 010000, Kazakhstan.
6
Department of Chemistry, School of Science and Technology, Nazarbayev University, Astana, 010000, Kazakhstan. Electronic address: darkhan.utepbergenov@nu.edu.kz.

Abstract

Mutations in the human protein DJ-1 cause early onset of Parkinson's disease. A reactive cysteine residue (Cys106) 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 Cys106 is carboxymethylated in a fraction of DJ-1 tightly bound to Ni-NTA and that the replacement of Cys106 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 Cys106 that transforms into a Cys106-carboxymethyl adduct upon elution from Ni-NTA. We conclude that an unknown metabolite reacts with Cys106 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.

KEYWORDS:

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

PMID:
30190129
DOI:
10.1016/j.bbrc.2018.08.190

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