Actin of chara giant internodal cells: a single isoform in the subcortical filament bundles and a larger, immunologically related protein in the chloroplasts

Internodal cells of Chara corallina Klein ex. Wild have been studied to determine the number of actin isoforms they contain and whether actin occurs at locations in the cortical cytoplasm outside the filament bundles. A monoclonal antibody to chicken actin is specific for actin in numerous animal cells but binds to two Chara proteins after their separation by two-dimensional polyacrylamide gel electrophoresis. One protein resembles known actins in relative molecular mass (43,000-M_r) and isoelectric point (5.5) while the other is distinctly different (58,000-M_r, isoelectric point = 4.8). Because it is indetectable in cells whose actin bundles have been extracted, the 43,000-M_r protein is assigned to the bundles and concluded to be rare or absent in the remaining cortical cytoplasm. The 58,000-M_r protein, in contrast, does not extract with the actin bundles. It was localized within the chloroplasts by immunofluorescence and by the dependence of proteolysis on the permeabilization of the chloroplast envelope.