A comparison of the intracellular distribution of mu-calpain, m-calpain, and calpastatin in proliferating human A431 cells.
Little is known about the relative intracellular localizations of the calcium-dependent proteases, calpains, and their naturally occurring inhibitor, calpastatin. In the present study, the intracellular localization of mu-calpain, m-calpain, and calpastatin was studied at the light microscopic level in proliferating A431 cells. Highly specific antibodies against the three antigens revealed distinct staining patterns in interphase and mitotic cells. Most notably, calpastatin in interphase cells was localized near the nucleus in tube-like, or large granular structures, while the calpains were more uniformly distributed through the cytoplasm in either a fibrillar form (mu-calpain) or a diffuse or fine granular form (m-calpain). The distribution patterns of the two calpain isozymes were distinctly different during mitosis. m-Calpain was concentrated at the mitotic spindle poles and midbody, while mu-calpain appeared to accumulate at the cell membrane and the spindles. Four other human cell lines as well as normal human monocytes were examined to determine if the calpains-calpastatin segregation patterns are common to other cells or are unique to the A431 line. With the exception of abundant nuclear mu-calpain in the C-33A cervical carcinoma, the segregation of the proteins was similar to that of A431. These studies indicate that calpains may be localized at regions which are relatively poor in calpastatin content. Proteins at these sites may be susceptible to calpain-catalyzed cleavage.
Lane
R D
RD
Department of Anatomy, Medical College of Ohio, Toledo 43699.
Allan
D M
DM
Mellgren
R L
RL
D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D002154:Calpain / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell; D002453:Cell Cycle / Q000502:physiology*; D002455:Cell Division / Q000502:physiology*; D003599:Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007399:Interphase; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; C033668:calpastatin; D002154:Calpain