Simple and rapid identification of phosphorylated peptides from bovine brain myelin basic protein by reversed-phase high-performance liquid chromatography.
The phosphorylation sites of the myelin basic protein from bovine brain were determined after phosphorylation with a cyclic 3':5'-phosphate-dependent protein kinase from the same source. Three phosphorylated peptides were selectively and rapidly separated, before and after dephosphorylation, by reversed-phase high-performance liquid chromatography on a styrene 250 column under alkaline conditions. Partial sequencing of the peptides by automated Edman degradation revealed that the serine-115 residue located in the main encephalitogenic determinant of the protein was a phosphorylation site, in addition to the two phosphorylation sites established (threonine-34 and serine-55).
Shoji
S
S
Ohnishi
J
J
Funakoshi
T
T
Kubota
Y
Y
Fukunaga
K
K
Miyamoto
E
E
Ueki
H
H
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006868:Hydrolysis; D004676:Myelin Basic Protein / Q000032:analysis*; D010455:Peptides / Q000032:analysis*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D014357:Trypsin
D000596:Amino Acids; D004676:Myelin Basic Protein; D010455:Peptides; D011494:Protein Kinases; D014357:Trypsin