Assessment of Proteolysis by Pyrylium and Other Fluorogenic Reagents

Author(s): Mohamed K. Al-Essa*, Ebaa Alzayadneh, Kamal Al-Hadidi

Journal Name: Protein & Peptide Letters

Volume 28 , Issue 7 , 2021


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Graphical Abstract:


Abstract:

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.

Keywords: Proteolysis, spectrofluorometry, pyrylium, CBQCA, chromeo P503, nanoOrange.

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Article Details

VOLUME: 28
ISSUE: 7
Year: 2021
Published on: 31 December, 2020
Page: [809 - 816]
Pages: 8
DOI: 10.2174/0929866528999201231214954

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