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A fluorescent assay of proteinases in cultured mammalian cells

F Dolbeare and M Vanderlaan

We have demonstrated proteinase activity in unfixed cells grown on tissue culture plates with a technique using 5-nitrosalicylaldehyde and peptide derivatives of 4-methoxy-2-naphthylamine. The 4-methoxy-2- naphthylamine liberated by proteinase activity reacts with 5- nitrosalicylaldehyde to form a fluorescent product. The substrates CBZ- alanyl-arginyl-arginyl-4-methoxy-2-naphthylamine and lysyl-alanyl-4- methoxy-2-naphthylamine, were used for the direct visual detection of two arylamidase activities in BALB/c 3T3 and C3H 10T 1/2 cells. With low magnification these enzyme activities can be detected in single clones; with higher magnification the fluorescent product can be seen within the cytoplasm of single cells.

Volume 27, Issue 11, pp. 1493-1495, 11/01/1979
Copyright © 1979 by The Histochemical Society


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