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

Journal Article · Sat Sep 15 04:00:00 EDT 2018 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/J.BBRC.2018.08.190· OSTI ID:23103551

Mussakhmetov, Arman ^[1]; Shumilin, Igor A. ^[2]; Nugmanova, Raushan ^[1]; Shabalin, Ivan G. ^[2]; Baizhumanov, Timur ^[3]; Toibazar, Daulet ^[4]; Khassenov, Bekbolat ^[1]; Minor, Wladek ^[2]; Utepbergenov, Darkhan ^[3]

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1. National Center for Biotechnology, Astana, 010000 (Kazakhstan)
2. Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, 22908 (United States)
3. Department of Chemistry, School of Science and Technology, Nazarbayev University, Astana, 010000 (Kazakhstan)
4. Department of Biology, School of Science and Technology, Nazarbayev University, Astana, 010000 (Kazakhstan)

Highlights: • A fraction of bacterially expressed polyhistidine-tagged human DJ-1 tightly binds to Ni-nitrilotriacetate (Ni-NTA) column. • The color of Ni-NTA column changes from light cyan to blue violet upon tight binding to DJ-1. • An active site cysteine is post-translationally modified in the fraction of DJ-1 tightly bound to Ni-NTA. • The modificationconferring tightbinding to Ni-NTA is unstable. • Crystal structure shows the unstable modification converts to S-carboxymethylcysteine upon elution from Ni-NTA. Mutations in the human protein DJ-1 cause early onset of Parkinson's disease. A reactive cysteine residue (Cys{sup 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{sup 106} is carboxymethylated in a fraction of DJ-1 tightly bound to Ni-NTA and that the replacement of Cys{sup 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{sup 106} that transforms into a Cys{sup 106}-carboxymethyl adduct upon elution from Ni-NTA. We conclude that an unknown metabolite reacts with Cys{sup 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.

• OSTI ID: 23103551
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 1 Vol. 504; ISSN 0006-291X; ISSN BBRCA9
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
COLOR
CRYSTAL STRUCTURE
IMIDAZOLES
NTA
SERINE