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 journalArticle

1 Citation (Scopus)

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 - 2000

Fingerprint

DEET
Pyridostigmine Bromide
Tubulin
Brain
Microtubules
Organophosphates
Colchicine
Conformations
Gas Poisoning
Insect Repellents
Microtubule Proteins
Cholinesterase Inhibitors
Protein Subunits
Stabilization

Keywords

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

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry

Cite this

Prasad, V., Scotch, R., Chaudhuri, A. R., Walss, C., Fathy, D. B., Miller, C., & Ludueña, R. F. (2000). Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl-m-toluamide. Neurochemical Research, 25(1), 19-25. https://doi.org/10.1023/A:1007527129743

Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl-m-toluamide. / Prasad, Veena; Scotch, Rebekah; Chaudhuri, Asish Ray; Walss, Consuelo; Fathy, Dana B.; Miller, Claudia; Ludueña, Richard F.

In: Neurochemical Research, Vol. 25, No. 1, 2000, p. 19-25.

Research output: Contribution to journalArticle

Prasad, V, Scotch, R, Chaudhuri, AR, Walss, C, Fathy, DB, Miller, C & Ludueña, RF 2000, 'Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl-m-toluamide', Neurochemical Research, vol. 25, no. 1, pp. 19-25. https://doi.org/10.1023/A:1007527129743
Prasad, Veena ; Scotch, Rebekah ; Chaudhuri, Asish Ray ; Walss, Consuelo ; Fathy, Dana B. ; Miller, Claudia ; Ludueña, Richard F. / Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl-m-toluamide. In: Neurochemical Research. 2000 ; Vol. 25, No. 1. pp. 19-25.
@article{ca5d1e57be2c440da84f5c913760ef7f,
title = "Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl-m-toluamide",
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.",
keywords = "Microtubule, N,N'-diethyl-m- toluamide, Pyridostigmine, Pyridostigmine bromide, Tubulin",
author = "Veena Prasad and Rebekah Scotch and Chaudhuri, {Asish Ray} and Consuelo Walss and Fathy, {Dana B.} and Claudia Miller and Ludue{\~n}a, {Richard F.}",
year = "2000",
doi = "10.1023/A:1007527129743",
language = "English (US)",
volume = "25",
pages = "19--25",
journal = "Neurochemical Research",
issn = "0364-3190",
publisher = "Springer New York",
number = "1",

}

TY - JOUR

T1 - Interactions of bovine brain tubulin with pyridostigmine bromide and N,N'-diethyl-m-toluamide

AU - Prasad, Veena

AU - Scotch, Rebekah

AU - Chaudhuri, Asish Ray

AU - Walss, Consuelo

AU - Fathy, Dana B.

AU - Miller, Claudia

AU - Ludueña, Richard F.

PY - 2000

Y1 - 2000

N2 - 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.

AB - 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.

KW - Microtubule

KW - N,N'-diethyl-m- toluamide

KW - Pyridostigmine

KW - Pyridostigmine bromide

KW - Tubulin

UR - http://www.scopus.com/inward/record.url?scp=0033978976&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033978976&partnerID=8YFLogxK

U2 - 10.1023/A:1007527129743

DO - 10.1023/A:1007527129743

M3 - Article

C2 - 10685600

AN - SCOPUS:0033978976

VL - 25

SP - 19

EP - 25

JO - Neurochemical Research

JF - Neurochemical Research

SN - 0364-3190

IS - 1

ER -