A role for microtubules in sorting endocytic vesicles in rat hepatocytes

Jason S. Goltz, Allan W. Wolkoff, Phyllis M. Novikoff, Richard J. Stockert, Peter Satir

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

The vectorial nature of hepatocyte receptor-mediated endocytosis (RME) and its susceptibility to cytoskeletal disrupters has suggested that a polarized network of microtubules plays a vital role in directed movement during sorting. Using as markers a well-known ligand, asialoorosomucoid, and its receptor, we have isolated endocytic vesicles that bind directly to and interact with stabilized endogenous hepatocyte microtubules at specific times during a synchronous, experimentally initiated, single wave of RME. Both ligand- and receptor-containing vesicles copelleted with microtubules in the absence of ATP but did not pellet under similar conditions when microtubules were not polymerized. When 5 mM ATP was added to preparations of microtubule-bound vesicles, ligand-containing vesicles were released into the supernatant, while receptor-containing vesicles remained immobilized on the microtubules. Release of ligand-containing vesicles from microtubules was prevented by monensin treatment during the endocytic wave. Several proteins, including the microtubule motor protein cytoplasmic dynein, were present in these preparations and were released from microtubule pellets by ATP addition concomitantly with ligand. These results suggest that receptor domains within the endosome can be immobilized by attachment to microtubules so that, following monensin-sensitive dissociation of ligand from receptor, ligand-containing vesicles can be pulled along microtubules away from the receptor domains by a motor molecule, such as cytoplasmic dynein, thereby delineating sorting.

Original languageEnglish (US)
Pages (from-to)7026-7030
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number15
StatePublished - 1992

Fingerprint

Transport Vesicles
Microtubules
Hepatocytes
Ligands
Cytoplasmic Dyneins
Monensin
Adenosine Triphosphate
Endocytosis
Asialoglycoprotein Receptor
Microtubule Proteins
Endosomes

Keywords

  • Asialoglycoprotein
  • Cytoplasmic dynein
  • Receptor-mediated endocytosis
  • Vesicular transport

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

A role for microtubules in sorting endocytic vesicles in rat hepatocytes. / Goltz, Jason S.; Wolkoff, Allan W.; Novikoff, Phyllis M.; Stockert, Richard J.; Satir, Peter.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 89, No. 15, 1992, p. 7026-7030.

Research output: Contribution to journalArticle

Goltz, Jason S. ; Wolkoff, Allan W. ; Novikoff, Phyllis M. ; Stockert, Richard J. ; Satir, Peter. / A role for microtubules in sorting endocytic vesicles in rat hepatocytes. In: Proceedings of the National Academy of Sciences of the United States of America. 1992 ; Vol. 89, No. 15. pp. 7026-7030.
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