Department of Physiology and Biochemistry, School of Medicine, The University of Jordan, Amman 11942, Jordan.
Department of Pathology, Microbiology and Forensic Medicine, School of Medicine, The University of Jordan, Amman 11942, Jordan.
Protein Pept Lett. 2021;28(7):809-816. doi: 10.2174/0929866528999201231214954.
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.
我们旨在评估胺反应荧光试剂在估计酶蛋白水解中的潜在应用。
蛋白酶在调节生物体许多生理过程中起着重要作用。
使用蛋白质测定技术的试剂评估蛋白质降解。
我们使用 Chromeo P503（Py1 吡喃鎓化合物）和 CBQCA（3-(4-羧基苯甲酰基)喹啉-2-甲醛）作为胺反应试剂和 NanoOrange 作为非胺反应染料，在与胰蛋白酶孵育 30-60 分钟后，对样本进行了测定。
用胺反应试剂 Chromeo P503（Py1 吡喃鎓化合物）和 CBQCA（3-(4-羧基苯甲酰基)喹啉-2-甲醛）测定时，所有用胰蛋白酶制备的 BSA 样本的荧光强度（FI）均明显高于对照样本（反映蛋白水解）。然而，用非胺反应试剂 NanoOrange 测定的相同样本在含胰蛋白酶样本和对照样本之间没有显示出任何显著差异。
这些结果证实了利用胺反应荧光试剂进行高度敏感的蛋白质分析，可用于一般估计蛋白水解的可靠性。
