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

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1980-06

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Abstract

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.

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10.1083/jcb.85.3.587

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Willard, M, C Simon, C Baitinger, J Levine and P Skene (1980). Association of an axonally transported polypeptide (H) with 100-A filaments. Use of immunoaffinity electron microscope grids. The Journal of cell biology, 85(3). pp. 587–596. 10.1083/jcb.85.3.587 Retrieved from https://hdl.handle.net/10161/21967.

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