Assessment of Proteolysis by Pyrylium and Other Fluorogenic Reagents.

AIMS

We aim to evaluate the potential application of amine reactive fluorogenic reagents for estimating enzymatic proteolysis.

BACKGROUND

Proteolytic enzymes play important roles in regulating many physiological processes in living organisms.

OBJECTIVES

Assessment of protein degradation by using reagents for protein assay techniques.

METHODS

We have assayed samples at the start and after 30-60 minutes incubation with trypsin by Chromeo P503 (Py 1 pyrylium compound) and CBQCA (3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde) as amine reactive reagents and NanoOrange as non-amine reactive dye.

RESULTS

All BSA prepared samples with trypsin have shown significantly higher fluorescence intensity (FI) versus controls (which reflects proteolysis) when assayed by Chromeo P503 (Py 1 pyrylium compound) and CBQCA (3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde) as amine reactive reagents. However, same samples assayed with NanoOrange as non-amine reactive reagent did not show any significant variation between samples containing trypsin and controls.

CONCLUSION

These results are confirming reliability of highly sensitive protein assays utilizing amine reactive fluorogenic reagents for general estimation of proteolysis.