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Assessment of Proteolysis by Pyrylium and Other Fluorogenic Reagents

2021; Bentham Science Publishers; Volume: 28; Issue: 7 Linguagem: Inglês

10.2174/0929866528999201231214954

1875-5305

Mohamed K. Al‐Essa, Ebaa Alzayadneh, Kamal Al-Hadidi,

Ubiquitin and proteasome pathways

We aim to evaluate the potential application of amine reactive fluorogenic reagents for estimating enzymatic proteolysis.Proteolytic enzymes play important roles in regulating many physiological processes in living organisms.Assessment of protein degradation by using reagents for protein assay techniques.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.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.These results are confirming reliability of highly sensitive protein assays utilizing amine reactive fluorogenic reagents for general estimation of proteolysis.