By John R. Griffiths, Richard D. Unwin
- Covers all significant ameliorations, together with phosphorylation, glycosylation, acetylation, ubiquitination, sulfonation and and glycation
- Discussion of the chemistry in the back of every one amendment, in addition to key equipment and references
- Contributions from a number of the top researchers within the field
- A precious reference resource for all laboratories project proteomics, mass spectrometry and post-translational amendment research
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Extra info for Analysis of protein post-translational modifications by mass spectrometry
Sikorski and Roland S. 1 Introduction to Protein Phosphorylation Much of the activity in the cellular proteome is under the control of reversible protein phosphorylation. Phosphorylation-dependent signaling regulates differentiation of cells, triggers progression of the cell cycle, and controls metabolism, transcription, apoptosis, and cytoskeletal rearrangements. Signaling via reversible protein phosphorylation also plays a critical role in intracellular communication and immune response. Phosphorylation can function as a positive or negative switch, activating or inactivating enzymes.
Surprisingly more phosphopeptides demonstrated better ionization efficiency than their nonphosphorylated counterparts, and in almost 70% of the cases the ratio of P/NP was within a factor of two . Most of these peptides were nontryptic and several contained multiple basic residues, particularly arginine. In the case of very basic peptides, the ionization efficiency was strikingly charge state dependent. In contrast, a separate study examining 66 model peptides and their singly phosphorylated counterparts found that in nearly all cases the phosphopeptides had lower ionization efficiency than their nonphosphorylated counterparts .
31 arises from the half of the sample that was treated with alkaline phosphatase. It represents the total amount of the sequence present in that half of the sample (sum of phosphopeptide and nonphosphopeptide). 81 comes from the half of the sample that was not treated with phosphatase, and it thus represents the original amount of nonphosphorylated peptide. 3 Determination of absolute stoichiometry. (a) Schematic showing the generalized protocol. Any suitable label that provides an adequate mass shift may be used.