Functional proteomics approaches for the identification of transnitrosylase and denitrosylase targets.
Methods
; 62(2): 151-60, 2013 Aug 01.
Article
em En
| MEDLINE
| ID: mdl-23428400
ABSTRACT
Protein S-nitrosylation is a dynamic post-translational modification (PTM) of specific cysteines within a target protein. Both proteins and small molecules are known to regulate the attachment and removal of this PTM, and proteins exhibiting such a function are transnitrosylase or denitrosylase candidates. With the advent of the biotin switch technique coupled to high-throughput proteomics workflows, the identification and quantification of large numbers of S-nitrosylated proteins and peptides is now possible. Proper analysis and interpretation of high throughout and quantitative proteomics data will help identify specific transnitrosylase and denitrosylase target peptide sequences and contribute to an understanding of the function and regulation of specific S-nitrosylation events. Here we describe the application of a quantitative proteomics approach using isotope-coded affinity tags (ICAT) in the biotin switch approach for the identification of transnitrosylation and denitrosylation targets of thioredoxin 1, an enigmatic protein with both reported transnitrosylase and denitrosylase activities.
Palavras-chave
2DE; ACN; Acetonitrile; BB; BCA; BST; Bicinchoninic acid; Binding buffer; Biotin switch; Biotin-HPDP; Biotin-switch technique; CID; Collision-induced dissociation; Cys32 and Cys35 to Ser Trx1 mutant; DDA; Data-dependent analysis; ENOA; ESI; Electrospray ionization; FDR; False discovery rate; GAPDH; GSNO; Glyceraldehyde 3-phosphate dehydrogenase; Human α-enolase; ICAT; ICAT heavy reagent; ICAT light reagent; ICAT-H; ICAT-L; IEF; Isobaric tag for relative and absolute quantitation; Isoelectric focusing; Isotope-coded affinity tags; LB; Lysis buffer; MALDI; MMTS; MRM; MS/MS; Matrix-assisted laser desorption ionization; Methyl methanethiosulfonate; Multiple-reaction monitoring; N-[6-(Biotinamido)hexyl]-3'-(2'-pyridyldithio)-propionamide; NADPH; NB; Nicotinamide adenine dinucleotide phosphate; PTM; Post-translational modification; Proteomics; RB; Resuspension buffer; S-Nitrosylated cysteine; S-Nitrosylated thioredoxin 1; S-Nitrosylation buffer; S-nitrosoglutathione; S-nitrosylation; SILAC; SNO-Cys; SNO-Trx1; SNO-site identification; SNOSID; Stable isotope labeling with amino acids in cell culture; TFA; TMT; Tandem Mass Tags; Tandem mass spectrometry; Thioredoxin; Trifluoroacetic acid; Trx; Trx reductase; Trx(C32S/C35S); TrxR; Two-dimensional gel electrophoresis; iTRAQ
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Tiorredoxinas
/
S-Nitrosotióis
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article