A substrate for DPP IV
Gly-Pro-pNA (hydrochloride) is a chromogenic substrate that can be cleaved by the circulating enzyme, dipeptidyl peptidase IV (DPP IV). Enzyme activity can be quantified by colorimetric detection of free p-nitroanilide at 405 nm. This substrate can be used to screen for DPP IV inhibitors.[5].
Mare whey proteins show good DPP-IV inhibitory activity in vitro，have the able to inhibit DPP-IV from degrading Gly-Pro-pNA (hydrochloride) Gly-Pro-pNA (hydrochloride) [1]. The in vitro DPP-IV inhibitory activities of quinoa protein hydrolysates can determined using Gly-Pro-pNA (hydrochloride) as a substrate[2]. Gly-Pro-pNA (hydrochloride) can also be used to evaluate the activity of DPP-IV in human serum[3]. Demonstrating that DPP IV in diabetic patients is not very different from that in normal conditions[4]. Gly-pro p-nitroanilide hydrochloride was used as a substrate to detect DPPIV level in patients with Polycystic syndrome (PCOS) ，a deregulation of DPP4 serum levels could be an additional characteristic of the metabolic imbalances associated with PCOS[5]
Using Gly-Pro-pNA (hydrochloride) as substrate can assess the enzymatic activity and biochemical status of dipeptidyl peptidase IV (DPP IV)in patients with rheumatoid arthritis[6]
参考文献:
[1]. Song JJ, Wang Q, et,al. Identification of dipeptidyl peptidase-IV inhibitory peptides from mare whey protein hydrolysates. J Dairy Sci. 2017 Sep;100(9):6885-6894. doi: 10.3168/jds.2016-11828. Epub 2017 Jul 12. PMID: 28711271.
[2]. You H, Wu T, et,al. Preparation and identification of dipeptidyl peptidase IV inhibitory peptides from quinoa protein. Food Res Int. 2022 Jun;156:111176. doi: 10.1016/j.foodres.2022.111176. Epub 2022 Mar 19. PMID: 35651037.
[3]. Yeganeh F, Mousavi SMJ, et,al. Association of CD26/dipeptidyl peptidase IV mRNA level in peripheral blood mononuclear cells with disease activity and bone erosion in rheumatoid arthritis. Clin Rheumatol. 2018 Dec;37(12):3183-3190. doi: 10.1007/s10067-018-4268-y. Epub 2018 Aug 22. PMID: 30136129.
[4]. Sun AL, Deng JT, et,al. Dipeptidyl peptidase-IV is a potential molecular biomarker in diabetic kidney disease. Diab Vasc Dis Res. 2012 Oct;9(4):301-8. doi: 10.1177/1479164111434318. Epub 2012 Mar 2. PMID: 22388283.
[5]. Blauschmidt S, Greither T, et,al. Dipeptidyl peptidase 4 serum activity and concentration are increased in women with polycystic ovary syndrome. Clin Endocrinol (Oxf). 2017 Dec;87(6):741-747. doi: 10.1111/cen.13444. Epub 2017 Sep 13. PMID: 28799235.
[6]. Mavropoulos JC, Cuchacovich M, et,al. Anti-tumor necrosis factor-alpha therapy augments dipeptidyl peptidase IV activity and decreases autoantibodies to GRP78/BIP and phosphoglucose isomerase in patients with rheumatoid arthritis. J Rheumatol. 2005 Nov;32(11):2116-24. PMID: 16265688.
Gly-Pro-pNA（盐酸盐）是一种显色底物，可被循环酶二肽基肽酶 IV (DPP IV) 裂解。酶活性可以通过在 405 nm 处游离对硝基苯胺的比色检测来量化。该底物可用于筛选 DPP IV 抑制剂。[5]。
马乳清蛋白在体外表现出良好的DPP-IV抑制活性，能够抑制DPP-IV降解Gly-Pro-pNA (hydrochloride) Gly-Pro-pNA (hydrochloride) [1]。以 Gly-Pro-pNA（盐酸盐）为底物，可以测定藜麦蛋白水解物的体外 DPP-IV 抑制活性[2]。 Gly-Pro-pNA（盐酸盐）也可用于评价 DPP-IV 在人血清中的活性[3]。证明糖尿病患者的 DPP IV 与正常情况下的差异不大[4]。以甘氨酰对硝基苯胺盐酸盐为底物检测多囊综合征（PCOS）患者的DPPIV水平，DPP4血清水平失调可能是PCOS相关代谢失衡的另一个特征[5]< /sup>
以Gly-Pro-pNA（盐酸盐）为底物可评估类风湿性关节炎患者二肽基肽酶IV（DPP IV）的酶活性和生化状态[6]
| Cell experiment [1]: | |
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Cell lines |
Mare whey protein hydrolysates |
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Preparation Method |
20 uL of sample solution and 100 uL of substrate solution (H- Gly-Pro-pNA (hydrochloride), final concentration 0.5 mM) were added. Enzyme reaction was started by adding DPP-IV. The absorbance at 405 nm was monitored after the microplate was incubated at 37°C for 0.5 h. |
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Reaction Conditions |
Final concentration 0.5 mM ，37 °C for 0.5 h. |
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Applications |
The degradation rate of the substrate Gly-Pro-pNA (hydrochloride) of DPP-IV was slowed by the addition of Mare Whey proteins. |
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参考文献: [1]. Song JJ, Wang Q, et,al. Identification of dipeptidyl peptidase-IV inhibitory peptides from mare whey protein hydrolysates. J Dairy Sci. 2017 Sep;100(9):6885-6894. doi: 10.3168/jds.2016-11828. Epub 2017 Jul 12. PMID: 28711271. |
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