ProtTech Inc. Protein Phosphorylation Mapping

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Protein Phosphorylation Mapping

Mapping protein phosphorylation sites is still a challenging task since: i) in vivo phosphorylated samples often have low stoichiometry of phosphorylation; ii). a protein amount is often very limited; which make it difficult to achieve high sequence coverage; iii) phosphorylated peptides generally have poor MS signal due to the negative charges in a phosphate group.

To overcome these difficulties, we have implemented following techniques for phosphorylation mapping:

A high performance Immobilized Metal Ion Chorography (IMAC) is used to purify phosphopeptides from proteolytic digested peptide mix.
An Ultra sensitive Nano-LC-MS/MS is used to sequence the purified phosphopeptides purified by IMAC.
To increase the sequence coverage of proteolytic digestions, we may carry out different proteolytic digestions. The complementary nature of these digestions can significantly increase the sequence coverage of a mapping.