A transient post-translational modification of active site cysteine alters binding properties of the parkinsonism protein DJ-1
- National Center for Biotechnology, Astana, 010000 (Kazakhstan)
- Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, 22908 (United States)
- Department of Chemistry, School of Science and Technology, Nazarbayev University, Astana, 010000 (Kazakhstan)
- 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, Vol. 504, Issue 1; Other Information: Copyright (c) 2018 Elsevier Inc. All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
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