Metal-catalyzed oxidation of bovine neurofilaments in vitro

Juan C Troncoso, Anthony C. Costello, James H. Kim, Gail V W Johnson

Research output: Contribution to journalArticle

Abstract

Neurofilaments (NF) are important determinants of the shape and size of nerve cells. The oxidation of NF, relevant to aging, neurodegenerative disorders, and axonal (Wallerian) degeneration, has not been studied. In this investigation, we have combined biochemical and ultrastructural methods to study the metal-catalyzed oxidation (MCO) of bovine NF using an ascorbate/ Fe+3/O2 system. The oxidation of NF proteins was documented by increases in carbonyl content, which were time- and concentration-dependent. Polyacrylamide gel electrophoresis (PAGE) and immunoblot analyses revealed the fragmentation of oxidized NF proteins, predominantly NF-H and NF-M. Electron microscopy (EM) showed that oxidized NF formed dense aggregates and bundles of laterally aggregated filaments. Finally, we also demonstrated that oxidized NF proteins were more susceptible to calpain proteolysis. In view of the growing evidence supporting increased oxidative stress on the nervous system in aging and the report of Cu/Zn superoxide dismutase mutation in familial motor neuron disease, oxidative injury of NF may be relevant to cell atrophy and degeneration of nerve cells and to the formation of abnormal cytoskeletal structures.

Original languageEnglish (US)
Pages (from-to)891-899
Number of pages9
JournalFree Radical Biology and Medicine
Volume18
Issue number5
DOIs
StatePublished - 1995

Fingerprint

Neurofilament Proteins
Intermediate Filaments
Neurons
Metals
Oxidation
Aging of materials
Proteolysis
Calpain
Oxidative stress
Neurology
Electrophoresis
Electron microscopy
Wallerian Degeneration
Motor Neuron Disease
In Vitro Techniques
Neurodegenerative Diseases
Nervous System
Atrophy
Polyacrylamide Gel Electrophoresis
Electron Microscopy

Keywords

  • Cytoskeleton
  • Free radicals
  • Metal
  • Neurofilament
  • Oxidation
  • Proteolysis

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)
  • Clinical Biochemistry
  • Medicine(all)
  • Toxicology

Cite this

Metal-catalyzed oxidation of bovine neurofilaments in vitro. / Troncoso, Juan C; Costello, Anthony C.; Kim, James H.; Johnson, Gail V W.

In: Free Radical Biology and Medicine, Vol. 18, No. 5, 1995, p. 891-899.

Research output: Contribution to journalArticle

Troncoso, Juan C ; Costello, Anthony C. ; Kim, James H. ; Johnson, Gail V W. / Metal-catalyzed oxidation of bovine neurofilaments in vitro. In: Free Radical Biology and Medicine. 1995 ; Vol. 18, No. 5. pp. 891-899.
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