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Phosphopeptide enrichment on synthetic libraries and HeLa cells digest

ABSTRACT: 2 phosphopeptide enrichment methods (TiO2 and Ti-IMAC) were evaluated on synthetic libraries and HeLa cells digest. The resulting peptides were compared to the ones without enrichment with regards to the nature of the phosphorylation site (serine, threonine, tyrosine) and the characteristics of the peptide enriched. It was found that the enrichment methods are unbiased for these properties and that the result of the enrichment reflects the original condition of the sample.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Lucrece Matheron

LAB HEAD: Albert Heck

PROVIDER: PXD000759 | Pride | 2014-08-11

REPOSITORIES: Pride

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Characterization of biases in phosphopeptide enrichment by Ti(4+)-immobilized metal affinity chromatography and TiO2 using a massive synthetic library and human cell digests.

Matheron Lucrece L van den Toorn Henk H Heck Albert J R AJ Mohammed Shabaz S

Analytical chemistry 20140804 16

Outcomes of comparative evaluations of enrichment methods for phosphopeptides depend highly on the experimental protocols used, the operator, the source of the affinity matrix, and the samples analyzed. Here, we attempt such a comparative study exploring a very large synthetic library containing thousands of serine, threonine, and tyrosine phosphorylated peptides, being present in roughly equal abundance, along with their nonphosphorylated counterparts, and use an optimized protocol for enrichme ...[more]

PMID: 25068997

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