Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro

Ronald D. Vale, Bruce J. Schnapp, Tim Mitchison, Eric Steuer, Thomas S. Reese, Michael Sheetz

Research output: Contribution to journalArticle

267 Citations (Scopus)

Abstract

Single microtubules from squid axoplasm support bidirectional movement of organelles. We previously purified a microtubule translocator (kinesin) that moves latex beads in only one direction along microtubules. In this study, a polar array of microtubules assembled off of centrosomes in vitro was used to demonstrate that kinesin moves latex beads from the minus to the plus ends of microtubules, a direction that corresponds to anterograde transport in the axon. A crude solubilized fraction from squid axoplasm (S1a), however, generates bidirectional movement of beads along microtubules. Retrograde bead movement (1.4 μm/sec) is inhibited by N-ethylmaleimide and 20 μM vanadate while anterograde movement (0.6 μm/sec) is unaffected by these agents. Furthermore, a monoclonal antibody against kinesin, when coupled to Sepharose, removes the anterograde, but not the retrograde, bead translocator from S1a. These results indicate that there is a retrograde bead translocator which is pharmacologically and immunologically distinct from kinesin.

Original languageEnglish (US)
Pages (from-to)623-632
Number of pages10
JournalCell
Volume43
Issue number3 PART 2
DOIs
StatePublished - Jan 1 1985
Externally publishedYes

Fingerprint

Kinesin
Microtubules
Latex
Decapodiformes
Proteins
Microspheres
Ethylmaleimide
Vanadates
Sepharose
Centrosome
Monoclonal Antibodies
Organelles
Axons
Direction compound
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro. / Vale, Ronald D.; Schnapp, Bruce J.; Mitchison, Tim; Steuer, Eric; Reese, Thomas S.; Sheetz, Michael.

In: Cell, Vol. 43, No. 3 PART 2, 01.01.1985, p. 623-632.

Research output: Contribution to journalArticle

Vale, RD, Schnapp, BJ, Mitchison, T, Steuer, E, Reese, TS & Sheetz, M 1985, 'Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro', Cell, vol. 43, no. 3 PART 2, pp. 623-632. https://doi.org/10.1016/0092-8674(85)90234-X
Vale, Ronald D. ; Schnapp, Bruce J. ; Mitchison, Tim ; Steuer, Eric ; Reese, Thomas S. ; Sheetz, Michael. / Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro. In: Cell. 1985 ; Vol. 43, No. 3 PART 2. pp. 623-632.
@article{e6160f2751b54c399231dc9bb05b6cea,
title = "Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro",
abstract = "Single microtubules from squid axoplasm support bidirectional movement of organelles. We previously purified a microtubule translocator (kinesin) that moves latex beads in only one direction along microtubules. In this study, a polar array of microtubules assembled off of centrosomes in vitro was used to demonstrate that kinesin moves latex beads from the minus to the plus ends of microtubules, a direction that corresponds to anterograde transport in the axon. A crude solubilized fraction from squid axoplasm (S1a), however, generates bidirectional movement of beads along microtubules. Retrograde bead movement (1.4 μm/sec) is inhibited by N-ethylmaleimide and 20 μM vanadate while anterograde movement (0.6 μm/sec) is unaffected by these agents. Furthermore, a monoclonal antibody against kinesin, when coupled to Sepharose, removes the anterograde, but not the retrograde, bead translocator from S1a. These results indicate that there is a retrograde bead translocator which is pharmacologically and immunologically distinct from kinesin.",
author = "Vale, {Ronald D.} and Schnapp, {Bruce J.} and Tim Mitchison and Eric Steuer and Reese, {Thomas S.} and Michael Sheetz",
year = "1985",
month = "1",
day = "1",
doi = "10.1016/0092-8674(85)90234-X",
language = "English (US)",
volume = "43",
pages = "623--632",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "3 PART 2",

}

TY - JOUR

T1 - Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro

AU - Vale, Ronald D.

AU - Schnapp, Bruce J.

AU - Mitchison, Tim

AU - Steuer, Eric

AU - Reese, Thomas S.

AU - Sheetz, Michael

PY - 1985/1/1

Y1 - 1985/1/1

N2 - Single microtubules from squid axoplasm support bidirectional movement of organelles. We previously purified a microtubule translocator (kinesin) that moves latex beads in only one direction along microtubules. In this study, a polar array of microtubules assembled off of centrosomes in vitro was used to demonstrate that kinesin moves latex beads from the minus to the plus ends of microtubules, a direction that corresponds to anterograde transport in the axon. A crude solubilized fraction from squid axoplasm (S1a), however, generates bidirectional movement of beads along microtubules. Retrograde bead movement (1.4 μm/sec) is inhibited by N-ethylmaleimide and 20 μM vanadate while anterograde movement (0.6 μm/sec) is unaffected by these agents. Furthermore, a monoclonal antibody against kinesin, when coupled to Sepharose, removes the anterograde, but not the retrograde, bead translocator from S1a. These results indicate that there is a retrograde bead translocator which is pharmacologically and immunologically distinct from kinesin.

AB - Single microtubules from squid axoplasm support bidirectional movement of organelles. We previously purified a microtubule translocator (kinesin) that moves latex beads in only one direction along microtubules. In this study, a polar array of microtubules assembled off of centrosomes in vitro was used to demonstrate that kinesin moves latex beads from the minus to the plus ends of microtubules, a direction that corresponds to anterograde transport in the axon. A crude solubilized fraction from squid axoplasm (S1a), however, generates bidirectional movement of beads along microtubules. Retrograde bead movement (1.4 μm/sec) is inhibited by N-ethylmaleimide and 20 μM vanadate while anterograde movement (0.6 μm/sec) is unaffected by these agents. Furthermore, a monoclonal antibody against kinesin, when coupled to Sepharose, removes the anterograde, but not the retrograde, bead translocator from S1a. These results indicate that there is a retrograde bead translocator which is pharmacologically and immunologically distinct from kinesin.

UR - http://www.scopus.com/inward/record.url?scp=0022294771&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022294771&partnerID=8YFLogxK

U2 - 10.1016/0092-8674(85)90234-X

DO - 10.1016/0092-8674(85)90234-X

M3 - Article

VL - 43

SP - 623

EP - 632

JO - Cell

JF - Cell

SN - 0092-8674

IS - 3 PART 2

ER -