Anti-idiotypic antibodies that react with microtubule-associated proteins are present in the sera of rabbits immunized with synthetic peptides from tubulin's regulatory domain.
A fundamental question in microtubule research is how the interactions of tubulin subunits with microtubule-associated proteins (MAPs) are controlled. The answer should provide insight into the regulation of the cellular processes in which microtubules are implicated. Previous work demonstrated the interaction of MAPs with a 4-kDa C-terminal domain of tubulin alpha and beta subunits. Synthetic peptides from the variable region of the 4-kDa C-terminal moiety of tubulin subunits, alpha-(430-441) and beta-(422-434), bind to MAP-2 and to the MAP tau, and a preferential interaction of the beta peptide is observed. To define the regulatory significance of the substructure of the C-terminal tubulin domain, we produced rabbit antisera against these MAP-interacting peptides. We found that these antisera contained not only antibodies to the original synthetic peptides but also antibodies to MAPs. Here, we report that these antibodies, which react with MAP-1, MAP-2, and tau, appear to be a population of anti-idiotypic antibodies directed to the anti-peptide antibodies. They can inhibit MAP-induced tubulin assembly into microtubules in vitro, and the addition of MAPs overcomes the inhibition. The recognition by these anti-idiotypic antibodies of the tubulin-binding domain on MAPs provides unequivocal evidence that the tubulin region defined by the synthetic peptides is directly involved in the interaction with MAPs.
Rivas
C I
CI
Department of Biochemistry, Biophysics, and Genetics, University of Colorado Health Sciences Center, Denver 80262.
Vera
J C
JC
Maccioni
R B
RB
D000818:Animals; D001323:Autoantibodies / Q000032:analysis*; D007114:Immunization; D007130:Immunoglobulin Idiotypes / Q000276:immunology*; D008869:Microtubule-Associated Proteins / Q000276:immunology*; D011817:Rabbits; D014404:Tubulin / Q000276:immunology*
D001323:Autoantibodies; D007130:Immunoglobulin Idiotypes; D008869:Microtubule-Associated Proteins; D014404:Tubulin