Effects of ATP and cyclic AMP on the in vitro assembly and stability of mammalian brain microtubules

Felix Pariente, Veena Prasad, Richard Froilan Ludueña, Rafael Manso-Martínez

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The relevance of protein phosphorylation, transphosphorylation and binding phenomena in the kinetics of the ATP-induced assembly of cycle-purified microtubule protein from mammalian brain were studied. ATP was able to induce the polymerization of microtubules of normal appearance. However, the assembled structures, were unstable and microtubules depolymerized after achievement of a transitory maximum. Cyclic AMP reduced the amplitude of the polymerization maximum in a concentration-dependent manner, correlating with the stimulation of the endogenous phosphorylation reaction. When microtubule assembly was induced by GTP, in the presence of various concentrations of ATP, the slope of the depolymerization phase was found to depend on the concentration of ATP. Fluoride ion inhibited the endogenous phosphorylation reaction and reduced the disassembly rate, in a concentration-dependent manner. Evidence is also presented indicating that ATP did not bind to phosphocellulose-purified tubulin. These results further contribute to indicate that ATP and cyclic AMP, acting coordinately to control the phosphorylation extent of microtubule proteins are important factors to determine microtubule stability within the cell. Some implications of this mechanism for the regulation by cAMP of the initiation of DNA synthesis and mitosis are considered.

Original languageEnglish (US)
Pages (from-to)43-54
Number of pages12
JournalMolecular and Cellular Biochemistry
Volume74
Issue number1
DOIs
StatePublished - Mar 1987

Fingerprint

Microtubules
Cyclic AMP
Brain
Phosphorylation
Adenosine Triphosphate
Microtubule Proteins
Polymerization
Depolymerization
Tubulin
Guanosine Triphosphate
Fluorides
Mitosis
In Vitro Techniques
Ions
Kinetics
DNA
Proteins

Keywords

  • assembly/diassembly
  • ATP
  • cAMP
  • microtubules
  • phosphorylation
  • stability

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Clinical Biochemistry
  • Cell Biology

Cite this

Effects of ATP and cyclic AMP on the in vitro assembly and stability of mammalian brain microtubules. / Pariente, Felix; Prasad, Veena; Ludueña, Richard Froilan; Manso-Martínez, Rafael.

In: Molecular and Cellular Biochemistry, Vol. 74, No. 1, 03.1987, p. 43-54.

Research output: Contribution to journalArticle

Pariente, Felix ; Prasad, Veena ; Ludueña, Richard Froilan ; Manso-Martínez, Rafael. / Effects of ATP and cyclic AMP on the in vitro assembly and stability of mammalian brain microtubules. In: Molecular and Cellular Biochemistry. 1987 ; Vol. 74, No. 1. pp. 43-54.
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