Site-specific radical directed dissociation of peptides at phosphorylated residues.
Site-specific fragmentation of peptides at phosphorylated serine or threonine residues is demonstrated. This radical directed cleavage is accomplished by a two-step procedure. First the phosphate is replaced with naphthalenethiol using well established Michael Addition chemistry. Second, the modified peptide is electrosprayed and subjected to irradiation at 266 nm. Absorption at naphthalene causes homolytic cleavage of the connecting carbon-sulfur bond yielding a radical in the beta-position. Subsequent rearrangement cleaves the peptide backbone yielding a d-type fragment. This chemistry is generally applicable as demonstrated by experiments with several different peptides. Assignment of phosphorylation sites is greatly facilitated by this approach, particularly for peptides containing multiple serine or threonine residues.
Diedrich
Jolene K
JK
Department of Chemistry, University of California, Riverside, California 92521, USA.
Julian
Ryan R
RR
D005609:Free Radicals / Q000737:chemistry; D009281:Naphthalenes / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D010766:Phosphorylation; D010777:Photochemistry; D012694:Serine / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013912:Threonine / Q000737:chemistry
D005609:Free Radicals; D009281:Naphthalenes; D010455:Peptides; D013438:Sulfhydryl Compounds; D013912:Threonine; D012694:Serine