Synthesis and use of an aminoquinolinone derivative for the fluorometric determination of oxytocinase

1989 ◽  
Vol 54 (10) ◽  
pp. 2802-2808 ◽  
Author(s):  
Hana P. Mašková ◽  
George Kokotos ◽  
Chryssa Tzougraki ◽  
Tomislav Barth
Keyword(s):  
Organic Solvent ◽  
Human Serum ◽  
Enzyme Reaction ◽  
Natural Substrate ◽  
Fluorogenic Substrate ◽  
Chromogenic Substrates ◽  
Michaelis Constant ◽  
Fluorometric Determination ◽  
Optimum Ph

A new fluorogenic substrate for the determination of the activity of human serum oxytocinase-cystine aminopeptidase (EC 3.4.11.3), H-Cys(Bzl)-NH-Meq, has been synthesized. The affinity of H-Cys(Bzl)-NH-Meq to oxytocinase was by two orders higher than that of the usually employed chromogenic substrates. The Michaelis constant of oxytocinase for this substrate was within the range of the optimum pH (7.0–7.5) 2.3.10-6 mol l-1, i.e. in the region of the affinity of the natural substrate, oxytocin. The concentration of dimethylsulfoxide used for the solubilization of H-Cys(Byl)-NH-Meq (<0.4%) did not influence adversely the course of the enzyme reaction as in the case of chromogenic substrates, where the concentrations of the organic solvent exceeded 3%.

New fluorogenic substrates for the determination of post-proline endopeptidase activity

1990 ◽  
Vol 55 (4) ◽  
pp. 1112-1118 ◽  
Author(s):  
Chryssa Tzougraki ◽  
George Kokotos ◽  
Hana P. Mašková ◽  
Eva Anzenbacherová ◽  
Tomislav Barth
Keyword(s):  
Enzyme Activity ◽  
Organic Solvent ◽  
Fluorogenic Substrate ◽  
Michaelis Constant ◽  
Fluorogenic Substrates ◽  
Endopeptidase Activity ◽  
Optimum Ph ◽  
Adverse Influence

A new fluorogenic substrate for determination of the activity of post-proline endopeptidase (EC 3.4.21.26), Z-Cys(Bzl)-Pro-NH-Meq has been synthesized. Affinity of this substrate to the enzyme was significantly higher than that of the hitherto employed substrates. The Michaelis constant of the post-proline endopeptidase for Z-Cys(Bzl)-Pro-NH-Meq was at the optimum pH (7.0) 1.05 10-6 mol l-1. The concentration of dimethylsulfoxide used for the solubilization of Z-Cys(Bzl)-Pro-NH-Meq (<0.4%) was outside the limits of adverse influence of the organic solvent on the enzyme activity. The fluorogenic substrate Z-Gly-Pro-NH-Meq was also synthesized for comparison (Km = 10.35 10-6 mol l-1 at pH 7.0).

Fluorometric determination of paraoxon in human serum using a gold nanoparticle-immobilized organophosphorus hydrolase and coumarin 1 as a competitive inhibitor

2013 ◽  
Vol 181 (1-2) ◽  
pp. 239-248 ◽  
Author(s):  
Nahid Kamelipour ◽  
Afshin Mohsenifar ◽  
Meisam Tabatabaei ◽  
Tavoos Rahmani-Cherati ◽  
Kamyar Khoshnevisan ◽  
...  
Keyword(s):  
Human Serum ◽  
Gold Nanoparticle ◽  
Competitive Inhibitor ◽  
Organophosphorus Hydrolase ◽  
Fluorometric Determination ◽  
Coumarin 1

scholarly journals Development of a fluorogenic small substrate for dipeptidyl peptidase-4

2017 ◽  
Vol 13 ◽  
pp. 2690-2697 ◽  
Author(s):  
Futa Ogawa ◽  
Masanori Takeda ◽  
Kanae Miyanaga ◽  
Keita Tani ◽  
Ryuji Yamazawa ◽  
...  
Keyword(s):  
Enzymatic Hydrolysis ◽  
Organic Compounds ◽  
Reaction Medium ◽  
Dipeptidyl Peptidase ◽  
Fluorogenic Substrate ◽  
Fluorometric Determination ◽  
Dipeptidyl Peptidase 4 ◽  
Fluorogenic Probes ◽  
Hydrolysis Of

A series of aniline and m-phenylenediamine derivatives with electron-withdrawing 3,3,3-trifluoropropenyl substituents were synthesized as small and chemically stable fluorescent organic compounds. Their fluorescence performances were evaluated by converting 2,4-disubstituted aniline 1 to the non-fluorescent dipeptide analogue H-Gly-Pro-1 for the use as a fluorogenic substrate for dipeptidyl peptidase-4 (DPP-4). The progress of the enzymatic hydrolysis of H-Gly-Pro-1 with DPP-4 was monitored by fluorometric determination of 1 released into the reaction medium. The results suggest that 1 could be used as fluorophore in OFF–ON-type fluorogenic probes.

Synthesis of peptide templated copper nanoclusters for fluorometric determination of Fe(III) in human serum

2016 ◽  
Vol 183 (10) ◽  
pp. 2831-2836 ◽  
Author(s):  
Ting Tang ◽  
Jiang Ouyang ◽  
Lanshuang Hu ◽  
Linyan Guo ◽  
Minghui Yang ◽  
...  
Keyword(s):  
Human Serum ◽  
Fluorometric Determination ◽  
Copper Nanoclusters

scholarly journals Fluorometric Determination of Tryptophan in Human Serum Using Phenylglyoxal.

1993 ◽  
Vol 9 (1) ◽  
pp. 25-27 ◽  
Author(s):  
Eijiro KOJIMA ◽  
Masaaki KAI ◽  
Yosuke OHKURA
Keyword(s):  
Human Serum ◽  
Fluorometric Determination

scholarly journals Fluorogenic Substrate [Ala-Pro]2-Cresyl Violet But Not Ala-Pro-Rhodamine 110 Is Cleaved Specifically by DPPIV Activity: A Study in Living Jurkat Cells and CD26/DPPIV-transfected Jurkat Cells

2003 ◽  
Vol 51 (7) ◽  
pp. 959-968 ◽  
Author(s):  
Emil Boonacker ◽  
Sjoerd Elferink ◽  
Abdennasser Bardai ◽  
Bernard Fleischer ◽  
Cornelis J.F. Van Noorden
Keyword(s):  
Microscopic Analysis ◽  
Enzyme Reaction ◽  
Proteolytic Cleavage ◽  
Living Cells ◽  
Jurkat Cells ◽  
Cresyl Violet ◽  
Fluorogenic Substrate ◽  
The Many ◽  
Kinetics Of

Fluorogenic substrates [Ala-Pro]2-cresyl violet and Ala-Pro-rhodamine 110 have been tested for microscopic detection of protease activity of dipeptidyl peptidase IV (DPPIV) in living cells. DPPIV activity is one of the many functions of the multifunctional or moonlighting protein CD26/DPPIV. As a model we used Jurkat cells, which are T-cells that lack CD26/DPPIV expression, and CD26/DPPIV-transfected Jurkat cells. Ala-Pro-rhodamine 110 is not fluorescent, but after proteolytic cleavage rhodamine 110 fluoresces. [Ala-Pro]2-cresyl violet is fluorescent by itself but proteolytic cleavage into cresyl violet induces a shift to longer wavelengths. This phenomenon enables the simultaneous determination of local (intracellular) substrate and product concentrations, which is important for analysis of kinetics of the cleavage reaction. [Ala-Pro]2-cresyl violet, but not Ala-Pro-rhodamine 110, appeared to be specific for DPPIV. When microscopic analysis is performed on living cells during the first minutes of the enzyme reaction, DPPIV activity can be precisely localized in cells with the use of [Ala-Pro]2-cresyl violet. Fluorescent product is rapidly internalized into submembrane granules in transfected Jurkat cells and is redistributed intracellularly via internalization pathways that have been described for CD26/DPPIV. We conclude that [Ala-Pro]2-cresyl violet is a good fluorogenic substrate to localize DPPIV activity in living cells when the correct wavelengths are used for excitation and emission and images are captured in the early stages of the enzyme reaction.

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