کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
867506 909784 2012 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Monitoring of proteolytic enzyme activity using phase transition-based peptide arrays
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Monitoring of proteolytic enzyme activity using phase transition-based peptide arrays
چکیده انگلیسی

We have developed an assay using peptide arrays based on phase transition from the glass substrate to the liquid for monitoring quantitative protease activity in real-time. Peptide arrays were fabricated using a bifunctional cross-linker, N-[γ-maleimidobutyryloxy] sulfosuccinimide ester, and a substrate peptide containing two functional groups, cysteine and tetramethyl-6-carboxyrhodamine (TAMRA) on the C- and N-terminus, respectively. The phase transition-based peptide arrays were characterized by analyzing the substrate peptide cleaved from the solid substrate by matrix metalloproteinase-3 (MMP-3). We successfully used this assay to determine the quantitative proteolytic activity of MMP-3 in a dose-dependent manner. In addition, parameters including Michaelis constant (Km), maximum rate of enzymatic reaction (Vmax), and half maximal inhibitory concentration (IC50) were determined by analyzing the concentrations of substrate peptide cleaved by MMP-3. Therefore, this new assay has potential for the quantitative analysis of enzyme kinetics of protease and informs research developments in drug discovery utilizing kinetic studies.


► We characterized phase transition-based peptide arrays for real-time monitoring of protease activity.
► We analyzed MMP-3 activity in real-time.
► We determined kinetic parameters, such as Km, Vmax, and IC50.
► This novel assay using peptides arrays were useful for quantitative analyses of protease kinetics.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biosensors and Bioelectronics - Volume 36, Issue 1, June–July 2012, Pages 147–153
نویسندگان
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