Combining Protein-Based IMAC, Peptide-Based IMAC, and MudPIT for Efficient Phosphoproteomic Analysis
Journal of Proteome Research2008Vol. 7(3), pp. 1346–1351
Citations Over TimeTop 1% of 2008 papers
Abstract
Immobilized metal affinity chromatography (IMAC) is a common strategy used for the enrichment of phosphopeptides from digested protein mixtures. However, this strategy by itself is inefficient when analyzing complex protein mixtures. Here, we assess the effectiveness of using protein-based IMAC as a pre-enrichment step prior to peptide-based IMAC. Ultimately, we couple the two IMAC-based enrichments and MudPIT in a quantitative phosphoproteomic analysis of the epidermal growth factor pathway in mammalian cells identifying 4470 unique phosphopeptides containing 4729 phosphorylation sites.
Related Papers
- Affinity chromatography : methods and protocols(2000)
- → Tyrosine hydroxylase phosphorylation is under the control of serine 40(2023)7 cited
- → Light-dependent phosphorylation of the gamma subunit of cGMP-phophodiesterase (PDE6γ) at residue threonine 22 in intact photoreceptor neurons(2009)13 cited
- Purification of Lectin in Hemolymph of Sarcophaga peregrina Larvae by Immuno affinity Chromatography(1999)
- The Effect of O-GlcNAcylation on Phosphorylation of tau(2003)