A putative beta-tubulin phosphate-binding motif is involved in lateral microtubule protofilament interactions

Eur J Biochem. 1997 Sep 15;248(3):840-7. doi: 10.1111/j.1432-1033.1997.t01-1-00840.x.

Abstract

We have investigated the role of a putative GTP-binding beta-tubulin motif in microtubule polymerization. A peptide containing residues 126-142 of the beta-tubulin subunit (peptide G) was synthesised and an antibody against it raised. Peptide G prevents the binding of GTP to tubulin and also microtubule polymerization but not the formation of vinblastine-induced tubulin spirals, suggesting that it may prevent lateral but not longitudinal tubulin-tubulin interactions. The antibody to peptide G shows little reaction with the interphase microtubule network, mitotic spindles or midbody of cultured cells, whereas it clearly reacts with vinblastine-induced paracrystals. These results suggest that this putative phosphate-binding site present in beta-tubulin could be involved in the lateral tubulin-tubulin interactions along the microtubule structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies / immunology
  • Binding Sites
  • Binding, Competitive
  • Dimerization
  • Fluorescent Antibody Technique
  • Guanosine Triphosphate / metabolism
  • Microscopy, Immunoelectron
  • Microtubules / metabolism*
  • Mitosis
  • Molecular Sequence Data
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / immunology
  • Peptide Fragments / metabolism
  • Peptide Fragments / pharmacology
  • Phosphates / metabolism
  • Protein Binding
  • Tubulin / chemistry*
  • Tubulin / metabolism*
  • Vinblastine / metabolism

Substances

  • Antibodies
  • Peptide Fragments
  • Phosphates
  • Tubulin
  • Vinblastine
  • Guanosine Triphosphate