Changes in microtubule-associated protein MAP1B phosphorylation during rat brain development.
Microtubule-associated protein MAP1B from neonatal rat brain was separated on sodium dodecyl sulfate-containing polyacrylamide gels into two isoforms (high and low MAP1B), both of which were recognized by a panel of monoclonal and polyclonal antibodies against MAP1B. In addition, SMI31, a monoclonal antibody directed against phosphorylated epitopes of the neurofilament proteins, showed phosphatase-sensitive reactivity against the high isoform of MAP1B. The antigenic relationship between the phosphorylated isoform of MAP1B and neurofilaments was confirmed by the reactivity of SMI31 with the immunoprecipitated MAP1B protein. After dephosphorylation of MAP1B with alkaline phosphatase, the higher-molecular-weight isoform of MAP1B was no longer detectable with phosphate-insensitive anti-MAP1B antibodies, whereas there was a significant increase in the immunoreactivity of the lower-molecular-weight MAP1B isoform. These data suggest that the structural microheterogeneity of MAP1B is due to differences in phosphorylation. The two isoforms were present in all brain regions of the young rat. During brain development, the general decrease in MAP1B levels was accompanied by changes in the relative amount of the two isoforms. In particular, the phosphorylated isoform of MAP1B decreased dramatically to almost undetectable levels in adult brain. This conclusion was further supported by immunoblotting analysis that showed the disappearance of phosphorylated epitopes of MAP1B early during brain development. In addition, dephosphorylation experiments demonstrated the phosphatase sensitivity of the phosphorylated isoform throughout development.
Fischer
I
I
Department of Biochemistry, E. K. Shriver Center, Waltham, MA 02254.
Romano-Clarke
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G
D000469:Alkaline Phosphatase / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn / Q000254:growth & development / Q000378:metabolism; D000911:Antibodies, Monoclonal; D001921:Brain / Q000254:growth & development* / Q000378:metabolism; D000939:Epitopes; D015151:Immunoblotting; D007536:Isomerism; D008869:Microtubule-Associated Proteins / Q000145:classification / Q000378:metabolism*; D008970:Molecular Weight; D010766:Phosphorylation; D051381:Rats
D000911:Antibodies, Monoclonal; D000939:Epitopes; D008869:Microtubule-Associated Proteins; C085284:microtubule-associated protein 1B; D000469:Alkaline Phosphatase