Discrete subcellular localization of a cytoplasmic and a mitochondrial isozyme of creatine kinase in intestinal epithelial cells.
Two isozymes of creatine kinase have been purified differentially from mitochondrial and cytoplasmic subfractions of intestinal epithelial cells. These intestinal epithelial cell creatine kinases were indistinguishable from the cytoplasmic (B-CK) and mitochondrial (Mi-CK) creatine kinase isozymes of brain when compared by SDS-PAGE, cellulose polyacetate electrophoresis, and peptide mapping. In intestinal epithelial cells, immunolocalization of the Mi-CK isozyme indicates that it is associated with long, thin mitochondria, which are excluded from the brush border at the apical end of each cell. In contrast, immunolocalization of the B-CK isozyme indicates that it is concentrated distinctly in the brush border terminal web domain. Although absent from the microvilli, B-CK also is distributed diffusely throughout the cytoplasm. Terminal web localization of B-CK was maintained in glycerol-permeabilized cells and in isolated brush borders, indicating that B-CK binds to the brush border structure. The abundance and localization of the mitochondrial and cytoplasmic creatine kinase isozymes suggest that they are part of a system that temporally and/or spatially buffers dynamic energy requirements of intestinal epithelial cells.
Keller
T C
TC
Department of Biological Science, Florida State University, Tallahassee 32312-3050.
Gordon
P V
PV
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002645:Chickens; D003402:Creatine Kinase / Q000276:immunology / Q000302:isolation & purification*; D003593:Cytoplasm / Q000737:chemistry / Q000201:enzymology* / Q000648:ultrastructure; D004386:Duodenum / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification*; D008871:Microvilli / Q000201:enzymology / Q000648:ultrastructure; D008928:Mitochondria / Q000737:chemistry / Q000201:enzymology* / Q000648:ultrastructure; D010449:Peptide Mapping
D000906:Antibodies; D007527:Isoenzymes; D003402:Creatine Kinase