Determination of the sites of posttranslational modifications in the charge isomers of bovine myelin basic protein by capillary electrophoresis-mass spectroscopy.
The posttranslational modifications in each of the 18.5 kDa bovine myelin basic protein charge isomers C-1 to C-6 have been determined by the use of capillary electrophoresis-mass spectroscopy. The pattern of modifications is viewed as being unique to each charge isomer and is thought to reflect a specific placement and function for each isomer in the myelin membrane. Several of the sites of posttranslational phosphorylation were found to differ from a number of the reported sites that were phosphorylated in vitro by various kinases. These differences suggest that an extremely cautious approach be taken in identifying in vivo posttranslationally modified amino acid residues from residues that have been modified in vitro by various kinases. We have identified the following posttranslationally phosphorylated and deamidated, modified sites in the bovine MBP components C1-C6. C1 has no modification; C2 represents a deamidation of Gln 146; in C3, Thr 97 and Ser 164 are phosphorylated; in C4, Ser 54, Thr 97, and Ser 160 are phosphorylated; in C5 Ser 7, Ser 54, Thr 97, and Ser 164 are phosphorylated; and in C6, Ser 7, Ser 54, Thr 97, Ser 160, and Ser 164 are phosphorylated.
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Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109-1055, USA.
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Lubman
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D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002417:Cattle; D019075:Electrophoresis, Capillary; D006801:Humans; D066298:In Vitro Techniques; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D014357:Trypsin
D004676:Myelin Basic Protein; D010446:Peptide Fragments; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D014357:Trypsin