Labelling of tumour cells with a biotinylated inhibitor of a cell surface protease.
Our objective has been to prepare a biotinylated affinity probe for the active centre of a protease associated with the surface of tumour cells. We employed three model systems in which easily recognisable tumour cells containing the active protease were used as targets for the biotinylated affinity probe. These were: squamous cell carcinoma, leukaemia cells in muscle and outgrowths of prostate carcinoma cells grown in three dimensional collagen gels. The presence of the bound biotinylated affinity probe was demonstrated by its ability to bind Texas-red labelled streptavidin with the results that the tumour cells exhibited red fluorescence. This binding was shown to be competitive with 9-amino acridine, a compound known to bind to the active centre of the target protease. This technique depends upon the affinity of the active centre of an enzyme for a competitive inhibitor and therefore should be applicable to other enzyme systems employing suitable ligands for their active centres.
Steven
F S
FS
Department of Biochemistry and Molecular Biology, School of Biological Sciences, University of Manchester, UK.
Griffin
M M
MM
Williams
L A
LA
Clarke
N W
NW
Maier
H
H
D000345:Affinity Labels / Q000378:metabolism*; D000376:Agmatine / Q000031:analogs & derivatives*; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives*; D002294:Carcinoma, Squamous Cell / Q000201:enzymology / Q000473:pathology; D002462:Cell Membrane / Q000201:enzymology*; D010450:Endopeptidases / Q000302:isolation & purification*; D006801:Humans; D007942:Leukemia, Experimental / Q000201:enzymology / Q000473:pathology; D008297:Male; D009062:Mouth Neoplasms / Q000201:enzymology / Q000473:pathology; D009369:Neoplasms / Q000201:enzymology* / Q000473:pathology; D011471:Prostatic Neoplasms / Q000201:enzymology / Q000473:pathology; D011480:Protease Inhibitors / Q000378:metabolism*; D051381:Rats
D000345:Affinity Labels; D011480:Protease Inhibitors; D001710:Biotin; D000376:Agmatine; D010450:Endopeptidases