Association of an axonally transported polypeptide (H) with 100-A filaments. Use of immunoaffinity electron microscope grids.

Published

Journal Article

Polypeptide H (mol wt 195,000) is axonally transported in rabbit retinal ganglion cells at a velocity of 0.7--1.1 mm/d, i.e., in the most slowly moving of the five transport groups described in these neurons. To identify the organelle with which H is associated, we purified H, prepared antibodies directed against it, and adsorbed the antibodies onto Formvar-coated electron microscope grids. When the resulting "immuno-affinity grids" were incubated with extracts of spinal cord and then examined in the electron microscope, they contained as many as 100 times more 100-A filaments than did grids coated similarly with nonimmune IgG. The ability of the anti-H IgG to specifically adsorb filaments to grids was completely blocked by incubating the IgG with polypeptide H. The 100-A filaments adsorbed to anti-H immunoaffinity grids could be specifically decorated by incubating them with anti-H IgG. These observations demonstrate that H antigens (and most likely H itself) are associated with 100-A neurofilaments. In addition, they suggest that the use of immunoaffinity grids may be a useful approach for determining the organelle associations of polypeptides.

Full Text

Duke Authors

Cited Authors

  • Willard, M; Simon, C; Baitinger, C; Levine, J; Skene, P

Published Date

  • June 1980

Published In

Volume / Issue

  • 85 / 3

Start / End Page

  • 587 - 596

PubMed ID

  • 6156174

Pubmed Central ID

  • 6156174

International Standard Serial Number (ISSN)

  • 0021-9525

Digital Object Identifier (DOI)

  • 10.1083/jcb.85.3.587

Language

  • eng

Conference Location

  • United States