Localization of the type III isozyme of hexokinase at the nuclear periphery.
The distribution of the type III isozyme of hexokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1) in rat kidney, liver, spleen, lung, and brain was determined immunohistochemically, using a monoclonal antibody generated against the enzyme purified from rat Novikoff hepatoma. In all tissues, specific cell types exhibited intense staining at the nuclear periphery, as confirmed by analysis using confocal microscopy. Isolated nuclei from kidney or liver were devoid of detectable type III hexokinase, although the enzyme was found in the "soluble" fraction from kidney or liver homogenates; these results suggest that the type III isozyme is associated in a labile manner with the external surface of the nucleus, with this association being disrupted by conventional homogenization and nuclear isolation procedures. The nuclear localization of the type III isozyme contrasts with previously demonstrated association of the type I and II isozymes with mitochondria. The physiological significance of a nuclear localization for the type III isozyme remains unclear. However, it was noted that many of the cells exhibiting prominent nuclear staining for type III hexokinase are endothelial or epithelial cells, suggesting a possible relationship between nuclear type III hexokinase and transport functions which are prominent in such cells.
Preller
A
A
Department of Biochemistry, Michigan State University, East Lansing 48824.
Wilson
J E
JE
D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000201:enzymology; D002467:Cell Nucleus / Q000201:enzymology* / Q000648:ultrastructure; D005260:Female; D006593:Hexokinase / Q000032:analysis*; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000032:analysis*; D007672:Kidney Cortex / Q000201:enzymology; D007679:Kidney Medulla / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008114:Liver Neoplasms, Experimental / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D006593:Hexokinase