کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
10533032 961840 2013 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A new proteinase 3 substrate with improved selectivity over human neutrophil elastase
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
A new proteinase 3 substrate with improved selectivity over human neutrophil elastase
چکیده انگلیسی
We report the synthesis and enzymatic studies on a new proteinase 3 intermolecular quenched substrate with enhanced selectivity over neutrophil elastase. Using combinatorial chemistry methods, we were able to synthesize the hexapeptide library with the general formula ABZ-Tyr-Tyr-Abu-X1′-X2′-X3′-Tyr(3-NO2)-NH2 using the mix and split method. The iterative deconvolution of such a library allowed us to obtain the sequence ABZ-Tyr-Tyr-Abu-Asn-Glu-Pro-Tyr(3-NO2)-NH2 with a high specificity constant (kcat/KM = 1534 × 103 M−1 s−1) and superior selectivity over neutrophil elastase and other neutrophil-derived serine proteases. Moreover, using the obtained substrate, we were able to detect a picomolar concentration of proteinase 3 (PR3). Incubation of the above-mentioned substrate with neutrophil lysate resulted in a strong fluorescent signal that was significantly reduced in the presence of a PR3 selective inhibitor.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 442, Issue 1, 1 November 2013, Pages 75-82
نویسندگان
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