Stable Isotope N-Phosphorylation Labeling for Peptide de Novo Sequencing and Protein Quantification Based on Organic Phosphorus Chemistry
2012
In this paper, we describe the development of a novel stable isotope N-phosphorylation labeling (SIPL) strategy for peptide de novo sequencing and protein quantification based on organic phosphorus chemistry. The labeling reaction could be performed easily and completed within 40 min in a one-pot reaction without additional cleanup procedures. It was found that N-phosphorylation labeling reagents were activated in situ to form labeling intermediates with high reactivity targeting on N-terminus and e-amino groups of lysine under mild reaction conditions. The introduction of N-terminal-labeled phosphoryl group not only improved the ionization efficiency of peptides and increased the protein sequence coverage for peptide mass fingerprints but also greatly enhanced the intensities of b ions, suppressed the internal fragments, and reduced the complexity of the tandem mass spectrometry (MS/MS) fragmentation patterns of peptides. By using nano liquid chromatography chip/time-of-flight mass spectrometry (nano LC-...
Keywords:
- Isobaric tag for relative and absolute quantitation
- Sample preparation in mass spectrometry
- Stable isotope labeling by amino acids in cell culture
- Analytical chemistry
- Isobaric labeling
- Terminal amine isotopic labeling of substrates
- Protein mass spectrometry
- Isotope-coded affinity tag
- Tandem mass tag
- Chemistry
- Biochemistry
- Chromatography
- Tandem mass spectrometry
- Mass spectrometry
- Correction
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