Evidence for a Ca2+-binding protein associated to non-actin microfilamentous systems in two ciliated protozoans

Electrophoretic distributions of proteins of isolated cortical cytoskeletons from two ciliated protozoans (Isotricha, Polyplastron) were compared in order to reveal any components common to non-actin microfilamentous structures. A low molecular weight protein (Mr approximately 22 kD) characterized by Ca2+-induced shifts in mobility on SDS-polyacrylamide gels was identified in both ciliates. Two-dimensional electrophoretic coordinates and peptide maps of Ca2+-binding proteins from Isotricha and Polyplastron are fairly similar, suggesting conservation of the same molecular species. In addition, an antiserum raised against two proteins (22-23 KD) from the filamentous ecto-endoplasmic boundary of Isotricha, one of which corresponds to the Ca2+-binding protein, cross-reacts specifically with that of Polyplastron. Using an immunogold staining procedure, the Ca2+-binding protein of Polyplastron was shown also to be located in a cortical microfilamentous layer. This protein is probably different from calmodulin. We postulate that it is involved in the control of the ordering of non-actin microfilaments within the cortex of ciliates.