Proteases that can distinguish among different post-translational forms of tyrosine engineered using multicolor flow cytometry.
Proteases that can distinguish among different post-translational forms of tyrosine engineered using multicolor flow cytometry.
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使用多色流式细胞术设计的蛋白酶可以区分酪氨酸的不同翻译后形式。
DOI：
10.1021/ja907803k
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发表时间：
2009
影响因子：
15
通讯作者：
Iverson,BrentL
中科院分区：
文献类型：
--
作者：
Varadarajan,Navin;Pogson,Mark;Georgiou,George;Iverson,BrentL
We report the isolation of a highly active (kcat/KM∼ 105M−1s−1) variant of theE. coliendopeptidase OmpT that selectively hydrolyzes peptides after 3-nitrotyrosine while effectively discriminating against similar peptides containing unmodified tyrosine (160-fold), sulfotyrosine (3600-fold), phosphotyrosine (>8000-fold), and phosphoserine (>8000-fold). The isolation of endopeptidase variants that can discriminate between substrates based on the post-translational modification of Tyr was made possible by implementing a multicolor flow cytometric assay for the screening of large mutant libraries. For the multicolor assay, a desired selection substrate was used simultaneously with multiple counterselection fluorescent substrates to isolate rare enzyme variants that displayed finely tuned substrate specificity. This work demonstrates that enzymes with exquisite selectivity can be isolated from large libraries using appropriate high throughput screening approaches and constitutes a critical step toward the production of a 3-nitrotyrosine-specific protease useful for proteomic applications.
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影响因子：
3.8
作者：
Woon-Seok Yeo;Soo Jae Lee;Jung Rok Lee;K. P. Kim
通讯作者：
Woon-Seok Yeo;Soo Jae Lee;Jung Rok Lee;K. P. Kim
利用大肠杆菌外膜内切蛋白酶 OmpT 变体作为加工酶从设计融合蛋白生产肽
影响因子：
4.4
作者：
K. Okuno;M. Yabuta;T. Ooi;S. Kinoshita
通讯作者：
S. Kinoshita
影响因子：
46.9
作者：
Santoro, SW;Wang, L;Schultz, PG
通讯作者：
Schultz, PG
影响因子：
14.8
作者：
Varadarajan, Navin;Rodriguez, Sarah;Iverson, Brent L.
通讯作者：
Iverson, Brent L.
一种工程蛋白酶，可特异性切割硫酸化酪氨酸。
DOI：
10.1002/anie.200800736
发表时间：
2008
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Varadarajan,Navin;Georgiou,George;Iverson,BrentL
通讯作者：
Iverson,BrentL