Title

Localization of an intermediate chain of outer arm dynein by immunoelectron microscopy

UMMS Affiliation

Department of Cell Biology

Date

11-15-1990

Document Type

Article

Subjects

Antibodies, Monoclonal; Centrifugation, Density Gradient; Chlamydomonas; Dynein ATPase; Immunoblotting; Immunosorbent Techniques; Macromolecular Substances; *Microscopy, Immunoelectron; Microtubules; Molecular Weight

Abstract

We have used immunoelectron microscopy to determine the location of an intermediate chain in the isolated outer arm dynein from Chlamydomonas flagella. When the purified alpha beta dimer of the outer arm was incubated with antibodies recognizing two distinct epitopes on its 69-kDa intermediate chain and then negatively stained and examined by electron microscopy, both antibodies appeared to have bound to the base of the Y-shaped stem that connects the two heads of the particle. These results indicate that this intermediate chain is located at the base of the stem. Inasmuch as this polypeptide is tightly associated with the 78-kDa intermediate chain and several light chains in an intermediate chain-light chain complex, it is likely that this entire assemblage is located at the base of the particle. Thus, these polypeptides are in a potentially important position with regard to the ATP-insensitive (structural end) binding of dynein to microtubules and to dynein-dynein interactions within the axoneme.

Rights and Permissions

Citation: J Biol Chem. 1990 Nov 15;265(32):19807-11.

Related Resources

Link to Article in PubMed

Journal Title

The Journal of biological chemistry

PubMed ID

2147183