Stable interaction between G-actin and neurofilament light subunit in dopaminergic neurons

R. Hao, R. G. Macdonald, M. Ebadi, J. C. Schmit, R. F. Pfeiffer

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Excessive accumulation of neurofilaments in the cell bodies and proximal axons of motor neurons is a major pathological hallmark of motor neuron diseases. In this communication we provide evidence that the neurofilament light subunit (68 kDa) and G-actin are capable of forming a stable interaction. Cytochalasin B, a cytoskeleton disrupting agent that interrupts actin-based microfilaments, caused aggregation of neurofilaments in cultured mesencephalic dopaminergic neurons, suggesting a possible interaction between neurofilaments and actin; which was tested further by using crosslinking reaction and affinity chromatography techniques. In the cross-linking experiment, G-actin interacted with individual neurofilament subunits and covalently cross-linked with disuccinimidyl suberate, a homobifunctional cross-linking reagent. Furthermore, G-actin was extensively cross-linked to the light neurofilament subunit with this reagent. The other two neurofilament subunits showed no cross-linking to G-actin. Moreover, neurofilament subunits were retained on a G-actin coupled affinity column and were eluted from this column by increasing salt concentration. All three neurofilament subunits became bound to the G-actin affinity column. However, a portion of the 160 and 200 kDa neurofilament subunits did not bind to the column, and the remainder of these two subunits eluted prior to the 68 kDa subunit, suggesting that the light subunit exhibited the highest affinity for G-actin. Moreover, neurofilaments demonstrated little or no binding to F-actin coupled affinity columns. The phosphorylation of neurofilament proteins with protein kinase C reduced its cross-linking to G-actin. The results of these studies are interpreted to suggest that the interaction between neurofilaments and actin, regulaled by neurofilament phosphorylation, may play a role in maintaining the structure and hence the function of dopaminergic neurons in culture.

Original languageEnglish (US)
Pages (from-to)825-834
Number of pages10
JournalNeurochemistry International
Volume31
Issue number6
DOIs
StatePublished - Dec 1 1997

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

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