Identification and localization of immunoreactive forms of caldesmon in smooth and nonmuscle cells: a comparison with the distributions of tropomyosin and alpha-actinin.
Caldesmon is an F-actin cross-linking protein of chicken gizzard smooth muscle whose F-actin binding activity can be regulated in vitro by Ca2+-calmodulin (Sobue, K., Y. Muramoto, M. Fujita, and S. Kakiuchi, 1981, Proc. Natl. Acad. Sci. USA, 78:5652-5655). It is a rod-shaped, heat-stable, F-actin bundling protein and is the most abundant F-actin cross-linking protein of chicken gizzard smooth muscle presently known (Bretscher, A., 1984, J. Biol. Chem., 259:12873-12880). We report the use of polyclonal antibodies to caldesmon to investigate its distribution and localization in other cells. Using immune blotting procedures, we have detected immunoreactive, heat-stable forms of caldesmon in cultured cells having either approximately the same apparent polypeptide molecular weight as gizzard caldesmon (120,000-140,000) or a substantially lower molecular weight (71,000-77,000). Through use of affinity-purified antibodies in indirect immunofluorescence microscopy, we have localized the immunoreactive forms to the terminal web of the brush border of intestinal epithelial cells and to the stress fibers and ruffling membranes of cultured cells. At the light microscope level caldesmon is distributed in a periodic fashion along stress fibers that is coincident with the distribution of tropomyosin and complementary to the distribution of alpha-actinin.
Bretscher
A
A
Lynch
W
W
D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D002148:Calmodulin-Binding Proteins; D002478:Cells, Cultured; D002645:Chickens; D005455:Fluorescent Antibody Technique; D006358:Hot Temperature; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism*; D008970:Molecular Weight; D009130:Muscle, Smooth / Q000378:metabolism / Q000648:ultrastructure*; D010749:Phosphoprotein Phosphatases / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D014335:Tropomyosin / Q000378:metabolism
D000199:Actins; D002148:Calmodulin-Binding Proteins; D008840:Microfilament Proteins; D014335:Tropomyosin; D000185:Actinin; D010749:Phosphoprotein Phosphatases