Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl-m-toluamide

Veena Prasad, Rebekah Scotch, Asish Ray Chaudhuri, Consuelo Walss, Dana B. Fathy, Claudia Miller, Richard F. Ludueña

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Pyridostigmine bromide (PB), an inhibitor of acetylcholinesterase, has been used as a prophylactic for nerve gas poisoning. N,N'-diethyl-m-toluamide (DEET) is the active ingredient in most insect repellents and is thought to interact synergistically with PB. Since PB can inhibit the binding of organophosphates to tubulin and since organophosphates inhibit microtubule assembly, we decided to examine the effects of PB and DEET on microtubule assembly as well as their interactions with tubulin, the subunit protein of microtubules. We found that PB binds to tubulin with an apparent K(d) of about 60 μM. PB also inhibits microtubule assembly in vitro, although at higher concentrations PB induces formation of tubulin aggregates of high absorbance. Like PB, DEET is a weak inhibitor of microtubule assembly and also induces formation of tubulin aggregates. Many tubulin ligands stabilize the conformation of tubulin as measured by exposure of sulfhydryl groups and hydrophobic areas and stabilization of colchicine binding. PB appears to have very little effect on tubulin conformation, and DEET appears to have no effect. Neither compound interferes with colchicine binding to tubulin. Our results raise the possibility that PB and DEET may exert some of their effects in vivo by interfering with microtubule assembly or function, although high intracellular levels of these compounds would be required.

Original languageEnglish (US)
Pages (from-to)19-25
Number of pages7
JournalNeurochemical Research
Volume25
Issue number1
DOIs
StatePublished - Feb 12 2000

Keywords

  • Microtubule
  • N,N'-diethyl-m- toluamide
  • Pyridostigmine
  • Pyridostigmine bromide
  • Tubulin

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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