Studies on the interaction and exchange of inhibitors and proteases on the surface of tumour cells in frozen sections.
We have used frozen sections of squamous cell carcinoma as a convenient source of a cell surface protease associated with tumour cells. This protease has been referred to as guanidinobenzoatase (GB) and is now known to be functionally identical to tissue plasminogen activator (t-PA). The use of a fluorescent competitive inhibitor of GB enabled the enzymic status of GB to be determined, i.e. was the enzyme active, latent or removed from our test system. The cell surface GB was then demonstrated to interact with extractable cytoplasmic inhibitors obtained from these sections. We then used a protected form of the GB in the absence of these internal inhibitors; such sections were used to transfer the GB to fibrin fibrils, thus exposing the presumptive receptor on the tumour cell surfaces. Texas red labelled t-PA was then shown to bind to the tumour cells in these pretreated sections from which the GB had previously been removed. We believe that the surface of tumour cells can be used to study the interaction of the naturally occurring inhibitors with GB and also that the cell surface receptors for GB can be used to study the binding of t-PA to cell surfaces.
Steven
F S
FS
Department of Biochemistry and Molecular Biology, School of Biological Sciences, University of Manchester, Manchester, UK.
Griffin
M M
MM
Bulleid
N J
NJ
Maier
H
H
Born
I A
IA
D000585:Aminacrine / Q000378:metabolism*; D001667:Binding, Competitive; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D003116:Color; D010450:Endopeptidases; D005337:Fibrin / Q000378:metabolism; D005629:Frozen Sections; D006801:Humans; D010959:Tissue Plasminogen Activator / Q000378:metabolism*; D014062:Tongue Neoplasms / Q000378:metabolism*
D000585:Aminacrine; D005337:Fibrin; D002265:Carboxylic Ester Hydrolases; C037288:guanidinobenzoate esterase; D010450:Endopeptidases; D010959:Tissue Plasminogen Activator