High-throughput characterization of intrinsic disorder in proteins from the Protein Structure Initiative.
High-throughput characterization of intrinsic disorder in proteins from the Protein Structure Initiative.
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DOI：
10.1016/j.jsb.2012.05.013
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发表时间：
2012-10
影响因子：
3
通讯作者：
Uversky, Vladimir N.
中科院分区：
文献类型：
--
作者：
Johnson, Derrick E.;Xue, Bin;Sickmeier, Megan D.;Meng, Jingwei;Cortese, Marc S.;Oldfield, Christopher J.;Le Gall, Tanguy;Dunker, A. Keith;Uversky, Vladimir N.
关键词：
在蛋白质结构计划中未能形成令人满意的晶体结构的目标中识别本质无序蛋白（IDP）是降低确定蛋白质三维结构的成本和时间的关键。为了帮助这一奋进，几个蛋白质结构倡议中心被要求发送可结晶蛋白质和未能结晶的蛋白质的样品。通过使用天然无序区预测因子（P0 NDR ®）的计算分析评估这些蛋白质中内在无序的丰度，并确定潜在的切割位点和相应的片段。然后，通过靶蛋白对有限蛋白水解的抗性来分析其内在紊乱。以溶菌酶/肌红蛋白、脱辅基肌红蛋白和α-酪蛋白为标准，分别作为有序、部分无序和完全无序蛋白，比较了胰蛋白酶对样品目标蛋白的消化速率。在下一阶段，蛋白质样品进行远紫外和近紫外圆二色性（CD）分析。对于大多数样本，CD数据、无序预测和有限胰蛋白酶消化率之间存在良好的一致性。正在对这些样品的一个较小的子集进行进一步的实验，以获得关于蛋白质有序/无序性质的更详细的信息。
The identification of intrinsically disordered proteins (IDPs) among the targets that fail to form satisfactory crystal structures in the Protein Structure Initiative represent a key to reducing the costs and time for determining three-dimensional structures of proteins. To help in this endeavor, several Protein Structure Initiative Centers were asked to send samples of both crystallizable proteins and proteins that failed to crystallize. The abundance of intrinsic disorder in these proteins was evaluated via computational analysis using Predictors of Natural Disordered Regions (PONDR®) and the potential cleavage sites and corresponding fragments were determined. Then, the target proteins were analyzed for intrinsic disorder by their resistance to limited proteolysis. The rates of tryptic digestion of sample target proteins were compared to those of lysozyme/myoglobin, apo-myoglobin and α-casein as standards of ordered, partially disordered and completely disordered proteins, respectively. At the next stage, the protein samples were subjected to both far-UV and near-UV circular dichroism (CD) analysis. For most of the samples, a good agreement between CD data, predictions of disorder and the rates of limited tryptic digestion was established. Further experimentation is being performed on a smaller subset of these samples in order to obtain more detailed information on the ordered/disordered nature of the proteins.
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影响因子：
6.8
作者：
Dunker, A. Keith;Silman, Israel;Sussman, Joel L.
通讯作者：
Sussman, Joel L.
影响因子：
2.9
作者：
Dunker, AK;Lawson, JD;Obradovic, Z
通讯作者：
Obradovic, Z
DOI：
10.1093/protein/11.5.349
发表时间：
1998-05-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Hubbard, SJ;Beynon, RJ;Thornton, JM
通讯作者：
Thornton, JM
影响因子：
5.8
作者：
Chen, L;Oughtred, R;Westbrook, J
通讯作者：
Westbrook, J
DOI：
10.1038/80747
发表时间：
2000-11-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Brenner, SE
通讯作者：
Brenner, SE