Computational design of an α-gliadin peptidase.
Computational design of an α-gliadin peptidase.
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DOI：
10.1021/ja3094795
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发表时间：
2012-12-19
影响因子：
15
通讯作者：
Siegel, Justin B.
中科院分区：
文献类型：
--
作者：
Gordon, Sydney R.;Stanley, Elizabeth J.;Wolf, Sarah;Toland, Angus;Wu, Sean J.;Hadidi, Daniel;Mills, Jeremy H.;Baker, David;Pultz, Ingrid Swanson;Siegel, Justin B.
合理修饰酶以进行新型化学转化的能力对于快速生产下一代蛋白质治疗剂至关重要。在这里，我们描述了使用化学原理来识别天然存在的酸活性肽酶，以及随后使用计算蛋白质设计工具来重新设计其对麸质中发现的免疫原性元素的特异性，麸质是乳糜泻的拟议原因。工程化的酶表现出的kcat/KM为568 M-1 s-1，代表了116倍以上的蛋白水解活性的模型面筋四肽比天然模板酶，以及超过800倍的开关底物特异性对面筋肽的免疫原性部分。计算工程酶对消化蛋白酶的蛋白水解具有抗性，并且在不到一小时内降解超过95%的与乳糜泻有关的免疫原性肽。因此，通过鉴定具有与最终目标相关的预先存在的品质的天然酶，并使用计算建模重新定义其底物特异性，我们能够产生具有作为乳糜泻治疗剂的潜力的酶。
The ability to rationally modify enzymes to perform novel chemical transformations is essential for the rapid production of next-generation protein therapeutics. Here we describe the use of chemical principles to identify a naturally occurring acid-active peptidase, and the subsequent use of computational protein design tools to reengineer its specificity toward immunogenic elements found in gluten that are the proposed cause of celiac disease. The engineered enzyme exhibits a kcat/KM of 568 M–1 s–1, representing a 116-fold greater proteolytic activity for a model gluten tetrapeptide than the native template enzyme, as well as an over 800-fold switch in substrate specificity toward immunogenic portions of gluten peptides. The computationally engineered enzyme is resistant to proteolysis by digestive proteases and degrades over 95% of an immunogenic peptide implicated in celiac disease in under an hour. Thus, through identification of a natural enzyme with the pre-existing qualities relevant to an ultimate goal and redefinition of its substrate specificity using computational modeling, we were able to generate an enzyme with potential as a therapeutic for celiac disease.
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影响因子：
15.3
作者：
Arentz-Hansen, H;Korner, R;Molberg, O;Quarsten, H;Vader, W;Kooy, Y M;Lundin, K E;Koning, F;Roepstorff, P;Sollid, L M;McAdam, S N
通讯作者：
McAdam, S N
影响因子：
56.9
作者：
Shan, L;Molberg, O;Khosla, C
通讯作者：
Khosla, C
DOI：
10.1124/jpet.106.104315
发表时间：
2006-09-01
影响因子：
3.5
作者：
Gass, Jonathan;Vora, Harmit;Khosla, Chaitan
通讯作者：
Khosla, Chaitan
影响因子：
3.7
作者：
Richter F;Leaver-Fay A;Khare SD;Bjelic S;Baker D
通讯作者：
Baker D
影响因子：
5.3
作者：
Wieser, Herbert
通讯作者：
Wieser, Herbert