کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1225406 968215 2011 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Identification of carbonylated peptides by tandem mass spectrometry using a precursor ion-like scan in negative ion mode
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Identification of carbonylated peptides by tandem mass spectrometry using a precursor ion-like scan in negative ion mode
چکیده انگلیسی

Reactive oxygen species (ROS) can oxidize proteins at almost any amino acid residue. Whereas some modifications are reversible within the cells, the higher oxidation states are especially irreversible. These irreversible post translational modifications are widely used as biomarkers of oxidative stress, such as protein carbonylation, which refers to aldehydes, ketones and lactams as ‘reactive carbonyl groups’. This study relied on a set of synthetic peptides containing a C-terminal aldehyde (arginal) or modification with pyruvic acid (ketone) or 4-hydroxynonenal (aldehyde) at lysine or histidine residues, as well as peptides containing pyroglutamic acid (oxidation product of proline) and 2-amino-3-butyric acid (oxidation product of threonine). The carbonylation sites were specifically derivatized with 2,4-dinitrophenylhydrazine (DNPH) and the fragmentation behavior of the products investigated in electrospray ionization (ESI-) MS. Importantly, the DNPH-labeled carbonylated peptides showed favorable ionization behaviors in negative ion mode ESI, providing a sensitive detection method. Regular peptides were mostly discriminated under these conditions. Among the fragmentation techniques tested for the negatively charged ions, pulsed Q dissociation provided three diagnostic ions at m/z values 152.0, 163.1 and 179.0, specific for DNPH-modified peptides. These marker ions were successfully applied to detect the carbonylated model peptides in a spiked tryptic digest of bovine serum albumin and a complex protein mixture obtained from HeLa cells.

Figure optionsDownload high-quality image (38 K)Download as PowerPoint slideHighlights
► Fragmentation behavior of DNPH derivatized carbonylated peptides was studied in negative ion mode.
► Three specific fragment ions at m/z 152.0, 163.1 and 179.0 were identified.
► A carbonylated peptide (30 pg) was successfully detected when spiked with a tryptic digest (150 ng) from HeLa cells.
► We conclude that ESI-MS in negative ion mode allows the sensitive detection of DNPH-carbonylated peptides.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Proteomics - Volume 74, Issue 11, 19 October 2011, Pages 2351–2359
نویسندگان
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