Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration

Nobutaka Hirokawa, K. Kevin Pfister, Hiroshi Yorifuji, Mark Wagner, Scott T. Brady, George S. Bloom

Research output: Contribution to journalArticle

329 Citations (Scopus)

Abstract

Kinesin is a microtubule-activated ATPase thought to transport membrane-bounded organelles along MTs. To illuminate the structural basis for this function, EM was used to locate submolecular domains on bovine brain kinesin. Rotary shadowed kinesin appeared rod-shaped and ∼80 nm long. One end of each molecule contained a pair of ∼10 × 9 nm globular domains, while the opposite end was fan-shaped. Monoclonal antibodies against the ∼124 kd heavy chains of kinesin decorated the globular structures, while those specific for the ∼64 kd light chains labeled the fanshaped end. Quick-freeze, deep-etch EM was used to analyze MTs polymerized from tubulin and cross-linked to latex microspheres by kinesin. Microspheres frequently attached to MTs by arm-like structures, 25-30 nm long. The MT attachment sites often appeared as one or two ∼10 nm globular buiges. Morphologically similar cross-links were observed by quick-freeze, deep-etch EM between organelles and MTs in the neuronal cytoskeleton in vivo. These collective observations suggest that bovine brain kinesin binds to MTs by globular domains that contain the heavy chains, and that the attachment sites for organelles are at the opposite, fan-shaped end of kinesin, where the light chains are located.

Original languageEnglish (US)
Pages (from-to)867-878
Number of pages12
JournalCell
Volume56
Issue number5
DOIs
StatePublished - Mar 10 1989
Externally publishedYes

Fingerprint

Kinesin
Electron microscopy
Brain
Electron Microscopy
Monoclonal Antibodies
Organelles
Microspheres
Fans
Light
Latex
Tubulin
Cytoskeleton
Microtubules
Adenosine Triphosphatases
Membranes
Molecules

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration. / Hirokawa, Nobutaka; Pfister, K. Kevin; Yorifuji, Hiroshi; Wagner, Mark; Brady, Scott T.; Bloom, George S.

In: Cell, Vol. 56, No. 5, 10.03.1989, p. 867-878.

Research output: Contribution to journalArticle

Hirokawa, Nobutaka ; Pfister, K. Kevin ; Yorifuji, Hiroshi ; Wagner, Mark ; Brady, Scott T. ; Bloom, George S. / Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration. In: Cell. 1989 ; Vol. 56, No. 5. pp. 867-878.
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