A new method which investigates lysosomal sensitivity to superoxide with endocytotic, site-specific, chemiluminescent probes.
The end result of cellular autophagocytosis and lysosomal digestion is incompletely understood in regard to the roles of the superoxide anion radical (O2-.). Cultured glial cells sequestered endocytotically two probes that were site-specific to the lysosome vacuome and sensitive to free-radical activities. The Sepharose-4B-polyisoluminol probe emitted chemiluminescent light in proportion to externally injected O2(-).. Bioluminescence measurements of beta-glucuronidase activity in intact glial cell lysosomes was achieved with the second site-specific, enzyme-specific Sepharose-4B-(dodecanate)5'-luciferinylglucuronate probe. Considerable activity was found in the lysosomal vacuome. In conclusion, we suggest that the use of site-specific, chemiluminescent probes can be of importance in the study of free radicals.
D052160:Benzothiazoles; D002478:Cells, Cultured; D004705:Endocytosis; D005609:Free Radicals; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D008163:Luminescent Measurements; D008165:Luminol / Q000031:analogs & derivatives; D008247:Lysosomes / Q000187:drug effects* / Q000502:physiology; D009457:Neuroglia / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology*; D011134:Polysaccharides; D011724:Pyridazines; D012685:Sepharose / Q000031:analogs & derivatives; D013481:Superoxides / Q000494:pharmacology*; D013844:Thiazoles
D052160:Benzothiazoles; D005609:Free Radicals; D007202:Indicators and Reagents; D011134:Polysaccharides; D011724:Pyridazines; D013844:Thiazoles; D013481:Superoxides; D008165:Luminol; C014525:sepharose-4B-(dodecanate)-5'-luciferinylglucuronate; C027239:sepharose-4B-polyisoluminol; D012685:Sepharose; D005966:Glucuronidase; D010100:Oxygen