The caldesmon content of vertebrate smooth muscle

Caldesmon and tropomyosin can be selectively and quantitatively extracted from vascular and visceral smooth muscle following heat treatment; all other smooth muscle proteins are precipitated by this procedure. Estimates of the caldesmon/tropomyosin molar ratio in heat-extracts determined by SDS-PAGE densitometry are 1 caldesmon:5.1-5.3 tropomyosin for rabbit and sheep aorta, and 1 caldesmon:5.9 tropomyosin for rabbit stomach and chicken gizzard. If the assumption is made that tropomyosin serves as a true reference of thin-filament content in intact muscle, it follows that the relative caldesmon contents in the above tissues are similar to each other. Caldesmon in heat extracts was identified by Western blotting, by its anomalous migration on several different SDS-PAGE systems and by its position on two-dimensional PAGE. Values of caldesmon contents in unfractionated total tissue homogenates were found to be similar to those cited above. Smooth muscles contain different thin-filament classes and only one type appears to possess caldesmon. By comparing values for the molar composition of caldesmon-specific filaments (1 caldesmon:2 tropomyosin:14 actin) with the values above determined for intact tissue, we conclude that the caldesmon filaments account for approx. 35-45% of the total thin-filament pool in arterial smooth muscle and slightly less in visceral muscles.