Quantitative identification of protein nitration sites.
Several labelling strategies have been developed targeting specific amino acid residues and/or PTMs. Methods specifically tailored for the qualitative and sometimes quantitative determination of PTMs have emerged. Many research groups have focused their attention towards o-nitrotyrosine residues, developing various methodologies for their identification, while direct quantification has remained elusive. So far the iTRAQ chemistry has been limited to primary amines. Here, we report a new strategy based on the use of iTRAQ reagents coupled to MS analysis for the selective labelling of o-nitrotyrosine residues. This method was proved to lead to the simultaneous localisation and quantification of nitration sites both in model proteins and in biological systems.
Chiappetta
Giovanni
G
Department of Organic Chemistry and Biochemistry, Federico II University of Naples, Naples, Italy.
Corbo
Claudia
C
Palmese
Angelo
A
Galli
Francesco
F
Piroddi
Marta
M
Marino
Gennaro
G
Amoresano
Angela
A
D000107:Acetylation; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins / Q000032:analysis / Q000737:chemistry; D008894:Milk Proteins / Q000032:analysis / Q000737:chemistry; D008969:Molecular Sequence Data; D009211:Myoglobin / Q000032:analysis / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
D000596:Amino Acids; D029968:Escherichia coli Proteins; D008894:Milk Proteins; D009211:Myoglobin; D010455:Peptides; D011506:Proteins; D012710:Serum Albumin, Bovine; C002744:3-nitrotyrosine; D014443:Tyrosine