Modification of the microtubule-binding and ATPase activities of kinesin by N-ethylmaleimide (NEM) suggests a role for sulfhydryls in fast axonal transport

K. Kevin Pfister, Mark Wagner, George S. Bloom, Scott T. Brady

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

N-Ethylmaleimide, an agent which alkylates free sulfhydryls in proteins, has been used to probe the role of sulfhydryls in kinesin, a motor protein for the movement of membrane-bounded organelles in fast axonal transport. When squid axoplasm is perfused with concentrations of NEM higher than 0.5 mM, organelle movements in both the anterograde and retrograde directions cease, and the vesicles remain attached to microtubules. Incubation of highly purified bovine brain kinesin with similar concentrations of NEM modifies the enzyme's microtubule-stimulated ATPase activity and promotes the binding of kinesin to microtubules in the presence of ATP. These results suggest that alkylation of sulfhydryls on kinesin alters the conformation of the protein in a manner that profoundly affects its interactions with ATP and microtubules. The NEM-sensitive sulfhydryls, therefore, may provide a valuable tool for the dissection of functional domains of the kinesin molecule and for understanding the mechanochemical cycle of this enzyme.

Original languageEnglish (US)
Pages (from-to)9006-9012
Number of pages7
JournalBiochemistry
Volume28
Issue number23
StatePublished - 1989
Externally publishedYes

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Kinesin
Axonal Transport
Ethylmaleimide
Microtubules
Adenosine Triphosphatases
Organelles
Adenosine Triphosphate
Dissection
Decapodiformes
Protein Conformation
Proteins
Alkylation
Enzymes
Conformations
Brain
Membrane Proteins
Membranes
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Modification of the microtubule-binding and ATPase activities of kinesin by N-ethylmaleimide (NEM) suggests a role for sulfhydryls in fast axonal transport. / Pfister, K. Kevin; Wagner, Mark; Bloom, George S.; Brady, Scott T.

In: Biochemistry, Vol. 28, No. 23, 1989, p. 9006-9012.

Research output: Contribution to journalArticle

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