RESUMO
S-nitrosylation (SNO) is an important oxidative posttranslational modification in the regulation of cardiac mitochondria. SNO modification of several mitochondrial proteins has been associated with cardiac preconditioning and improved cell survival following ischemia/reperfusion injury. Due to their labile nature, SNO modifications are challenging to study using traditional biochemical techniques; particularly, the identification of individual modified cysteine residues. Here, we describe the details of the cysTMT(6) switch assay, a variation of the classic biotin switch protocol. The cysTMT(6) reagent provides a simplified and powerful approach to SNO detection by combining unambiguous identification of the modified cysteine residue and relative quantification of up to six samples by mass spectrometry analysis.
Assuntos
Bioensaio , Cisteína/química , Mitocôndrias Cardíacas/metabolismo , Proteínas Mitocondriais/metabolismo , Miocárdio/metabolismo , Processamento de Proteína Pós-Traducional , Animais , Biotina/química , Cisteína/metabolismo , Mitocôndrias Cardíacas/efeitos dos fármacos , Proteínas Mitocondriais/química , Óxido Nítrico/biossíntese , Oxirredução , Ratos , S-Nitrosoglutationa/farmacologiaRESUMO
Macrolide magic: An enyne cross-metathesis reaction of an alkynyl boronate with an alkene derivative as well as a radical cyclization reaction of a homopropargylic beta-alkoxyacrylate are the key transformations in the total synthesis of the cytotoxic macrolide (-)-amphidinolide K.
