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Assessment of Proteolysis by Pyrylium and Other Fluorogenic Reagents

Author(s):

Mohamed K. Al-Essa*, Ebaa Alzayadneh and Kamal Al-Hadidi   Pages 1 - 8 ( 8 )

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.

Affiliation:

Department of Physiology and Biochemistry, School of Medicine, The University of Jordan, Amman 11942,, Department of Physiology and Biochemistry, School of Medicine, The University of Jordan, Amman 11942,, Department of Pathology, Microbiology and Forensic Medicine, School of Medicine, The University of Jordan, Amman 11942



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