Determination of nitrated peptides from αSyn and A30P.

Abstract

<p>αSyn and A30P were enriched by Ni²⁺ pull-down from yeast crude extracts and separated by SDS-PAGE. Monomeric and dimeric αSyn stained with Coomassie were excised from the gel and digested with trypsin and AspN. Untreated <i>(in vivo)</i> and subsequent peroxynitrite (PON) treated <i>(in vitro)</i> αSyn and A30P protein samples were analyzed with LC-MS for tyrosine nitration. 3-NT (3-nitrotyrosine) indicates identified nitration sites, supported by at least two peptides and two independent experiments.</p