Electron microscopic studies of chicken gizzard caldesmon and its complex with calmodulin

Abstract

Caldesmon samples mounted on a stage rotating about a horizontal axis were shadowed keeping the shadow angle at about 3 degrees. This technique minimizes background metal deposits compared with the conventional method. The identity of caldesmon was confirmed by comparing the images of caldesmon alone with those of the caldesmon-calmodulin complex. In these samples the caldesmon molecules appeared to be elongated; most were between 30 and 80 nm in length. The maximum length was in good agreement with the earlier estimate of 74 nm based on hydrodynamic studies. Our observations also suggested the presence of a rather rigid 30-40 nm stretch in the middle of the caldesmon molecule, which was always visible under rotary shadowing, and a flexible structure of about 20 nm in length at each end of the molecule, which may or may not be visible depending on their orientation on the mica surface. In the samples of caldesmon crosslinked with calmodulin, we noticed the existence of complexes containing two calmodulin molecules per caldesmon molecule, separated by a distance of 60 nm, consistent with the suggestion that each end of caldesmon can interact with calmodulin.