Identification of phosphorylation sites in polypeptides by employment of uranyl photocleavage
AssigneeUniversity of Copenhagen
InventorsNiels Erik Mollegaard, Peter Eigil Nielsen
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Abstract
The present invention relates to a method of cleaving a polypeptide at one or more phosphorylated residues. Said cleavage is induced by irradiation and is dependent on the presence of uranyl. The method is useful for analysis of phosphoproteoms and also for protein purification. The method also relates to a method of protein purification, wherein the phosphorylated protein is immobilized on a column said immobilization being dependent on uranyl.
Claims59
A method of cleaving a polypeptide at one or more phosphorylated residues comprising the steps of:
- a. Providing a sample comprising a phosphorylated polypeptide;
- b. Providing a sample comprising uranyl;
- c. Adding the sample comprising uranyl to the sample comprising the phosphorylated polypeptide to provide a uranyl-polypeptide sample;
- and
- d. Irradiating the uranyl-polypeptide sample, and thereby photo cleaving the phosphorylated polypeptide at phosphorylated residues.
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A method of purifying a phosphorylated protein from a sample, wherein the phosphorylated protein is immobilized on a column via uranyl coordinated to phosphorylated sites on the protein.
Dependencies are inferred from the claim text.