Phosphorylation States of Actin Filament Adenine Nucleotides in Detergent-Extracted Neuronal Cytoskeletal Fractions

James M Angelastro, D. L. Purich

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

High performance liquid chromatography of nucleotides from Triton X-100 cytoskeletal extracts has permitted analysis of the ATP and ADP content of actin filaments isolated intact from PC12 pheochromocytoma cells. We observed that the adenine nucleotide content matched the actin content of these cytoskeletal extracts, a finding consistent with the unit stoichiometry of nucleotide binding. Efficient assembly-linked ATP hydrolysis occurs in vivo, and based on a boundary hydrolysis model for nucleotide-promoted assembly, the observed ADP/ATP ratio indicates that the average microfilament in nonmuscle cells has 24 ATP-actin molecules at its growing end. Studies with NB41A3 neuroblastoma cells indicate that the ATP content of assembled actin filaments is about 3-4 times lower.

Original languageEnglish (US)
Pages (from-to)1490-1494
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume201
Issue number3
DOIs
StatePublished - Jun 30 1994
Externally publishedYes

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Phosphorylation
Adenine Nucleotides
Actin Cytoskeleton
Detergents
Actins
Adenosine Triphosphate
Nucleotides
Hydrolysis
PC12 Cells
Octoxynol
High performance liquid chromatography
Pheochromocytoma
Neuroblastoma
Stoichiometry
Adenosine Diphosphate
High Pressure Liquid Chromatography
Molecules

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

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AB - High performance liquid chromatography of nucleotides from Triton X-100 cytoskeletal extracts has permitted analysis of the ATP and ADP content of actin filaments isolated intact from PC12 pheochromocytoma cells. We observed that the adenine nucleotide content matched the actin content of these cytoskeletal extracts, a finding consistent with the unit stoichiometry of nucleotide binding. Efficient assembly-linked ATP hydrolysis occurs in vivo, and based on a boundary hydrolysis model for nucleotide-promoted assembly, the observed ADP/ATP ratio indicates that the average microfilament in nonmuscle cells has 24 ATP-actin molecules at its growing end. Studies with NB41A3 neuroblastoma cells indicate that the ATP content of assembled actin filaments is about 3-4 times lower.

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