Phosphopeptide enrichment for phosphoproteomic analysis - A tutorial and review of novel materials
| Autor: | Trong Khoa Pham, Andrew Landels, Caroline A. Evans, Phillip C. Wright, Wen Qiu |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Phosphopeptides
Proteomics Phosphorylation sites Peptide Single step 02 engineering and technology Computational biology 01 natural sciences Biochemistry Chromatography Affinity Analytical Chemistry Affinity chromatography Environmental Chemistry Phosphorylation Spectroscopy Enhanced selectivity Titanium chemistry.chemical_classification Phosphopeptide Chemistry 010401 analytical chemistry Phosphoproteomics 021001 nanoscience & nanotechnology 0104 chemical sciences 0210 nano-technology Protein Processing Post-Translational |
| Zdroj: | Analytica Chimica Acta. 1129:158-180 |
| ISSN: | 0003-2670 |
| DOI: | 10.1016/j.aca.2020.04.053 |
| Popis: | Significant technical advancements in phosphopeptide enrichment have enabled the identification of thousands of p-peptides (mono and multiply phosphorylated) in a single experiment. However, it is still not possible to enrich all p-peptide species in a single step. A range of new techniques and materials has been developed, with the potential to provide a step-change in phosphopeptide enrichment. The first half of this review contains a tutorial for new potential phosphoproteomic researchers; discussing the key steps of a typical phosphoproteomic experiment used to investigate canonical phosphorylation sites (serine, threonine and tyrosine). The latter half then show-cases the latest developments in p-peptide enrichment including: i) Strategies to mitigate non-specific binding in immobilized metal ion affinity chromatography and metal oxide affinity chromatography protocols; ii) Techniques to separate multiply phosphorylated peptides from monophosphorylated peptides (including canonical from non-canonical phosphorylated peptides), or to simultaneously co-enrich other post-translational modifications; iii) New hybrid materials and methods directed towards enhanced selectivity and efficiency of metal-based enrichment; iv) Novel materials that hold promise for enhanced phosphotyrosine enrichment. A combination of well-understood techniques and materials is much more effective than any technique in isolation; but the field of phosphoproteomics currently requires benchmarking of novel materials against current methodologies to fully evaluate their utility in peptide based proteoform analysis. |
| Databáze: | OpenAIRE |
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