Synthesis of diazo functionalized solid supports and their application towards the enrichment of phosphorylated peptides.
We describe herein the synthesis and application of a range of diazo-functionalized materials as platforms for the solid phase enrichment of phosphorylated peptides from protein digests. Diazo-functionalized solid phase resins are employed to bind covalently and specifically with the phosphate moiety of a peptide substrate, allowing for non-phosphorylated peptides to be efficiently removed by filtration. The technique described herein is compatible with enrichment of phospho-serine, threonine and tyrosine residues and was used to successfully enrich phosphorylated substrates from beta-casein and ovalbumin digests in low-picomolar quantities.
Frawley Cass
Samantha M
SM
Department of Chemistry, Michigan State University, East Lansing, MI 48824, USA.
Reid
Gavin E
GE
Tepe
Jetze J
JJ
D002364:Caseins / Q000378:metabolism; D003979:Diazonium Compounds / Q000737:chemistry*; D005374:Filtration / Q000379:methods; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D010766:Phosphorylation; D013912:Threonine / Q000737:chemistry*; D014443:Tyrosine / Q000737:chemistry*
D002364:Caseins; D003979:Diazonium Compounds; D010455:Peptides; D013912:Threonine; D014443:Tyrosine