TY - JOUR
T1 - A transient post-translational modification of active site cysteine alters binding properties of the parkinsonism protein DJ-1
AU - Mussakhmetov, Arman
AU - Shumilin, Igor A.
AU - Nugmanova, Raushan
AU - Shabalin, Ivan G.
AU - Baizhumanov, Timur
AU - Toibazar, Daulet
AU - Khassenov, Bekbolat
AU - Minor, Wladek
AU - Utepbergenov, Darkhan
PY - 2018/9/26
Y1 - 2018/9/26
N2 - 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.
AB - 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.
KW - DJ-1
KW - Oxidative stress
KW - PARK7
KW - Parkinson's disease
KW - S-carboxymethylcysteine
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U2 - 10.1016/j.bbrc.2018.08.190
DO - 10.1016/j.bbrc.2018.08.190
M3 - Article
C2 - 30190129
AN - SCOPUS:85052817315
VL - 504
SP - 328
EP - 333
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -