The protective role of a natural inhibitor in the fluorescent location of cells possessing a latent form of cell surface protease.
Leukaemia cells possess a latent form of a cell surface protease referred to as guanidinobenzoatase. Latency is due to complex formation between an inhibitor protein and the cell surface enzyme which is stable under acid conditions but is dissociated with formaldehyde treatment. The latent form of the cell surface protease has been used as a protecting mechanism during a preliminary step to stain all the nuclei of cells with haematoxylin. The enzyme-inhibitor complex was then dissociated and a combination of 9-amino acridine and propidium iodide employed to enable the fluorescent location of cells possessing active guanidinobenzoatase. We were thus able to visualise the nuclei by conventional light microscopy and simultaneously visualise the cell surface of leukaemia cells by fluorescent microscopy. This simple model system has provided technology applicable to the more complex analysis of neoplastic cells in cervical smears.
Steven
F S
FS
Department of Biochemistry and Molecular Biology, School of Biological Sciences, University of Manchester, UK.
Williams
L A
LA
D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000032:analysis / Q000037:antagonists & inhibitors*; D002462:Cell Membrane / Q000201:enzymology* / Q000648:ultrastructure; D010450:Endopeptidases; D006801:Humans; D007938:Leukemia / Q000201:enzymology* / Q000473:pathology; D007942:Leukemia, Experimental / Q000201:enzymology* / Q000473:pathology; D008856:Microscopy, Fluorescence; D011480:Protease Inhibitors / Q000032:analysis*; D051381:Rats; D013194:Staining and Labeling
D011480:Protease Inhibitors; D002265:Carboxylic Ester Hydrolases; C037288:guanidinobenzoate esterase; D010450:Endopeptidases