DE NOVO PROTEIN STRUCTURE GENERATION FROM INCOMPLETE CHEMICAL SHIFT ASSIGNMENTS

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 NMR化学位移为蛋白质提供了重要的局部结构信息。一致的结构生成NMR化学位移数据最近已成为可行的蛋白质的大小高达130个残基，这样的结构是一个质量与标准的NMR协议获得的。本研究探讨了化学位移归属的完整性对化学位移产生的结构的影响。化学位移Rosetta（CS-Rosetta）方案用于从头蛋白质结构生成，具有不同程度的化学位移分配完整性，通过省略先前用于CS-Rosetta方法初始演示的实验化学位移数据中的条目进行模拟。此外，描述了一种新的CS-Rosetta协议，该协议提高了该方法对于具有丢失或错误的NMR化学位移输入数据的蛋白质的鲁棒性。这种策略，它使用传统的Rosetta预过滤的片段选择过程中，证明了两个顺磁性蛋白质，也为两个蛋白质与固态NMR化学位移分配。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. NMR chemical shifts provide important local structural information for proteins. Consistent structure generation from NMR chemical shift data has recently become feasible for proteins with sizes of up to 130 residues, and such structures are of a quality comparable to those obtained with the standard NMR protocol. This study investigates the influence of the completeness of chemical shift assignments on structures generated from chemical shifts. The Chemical-Shift-Rosetta (CS-Rosetta) protocol was used for de novo protein structure generation with various degrees of completeness of the chemical shift assignment, simulated by omission of entries in the experimental chemical shift data previously used for the initial demonstration of the CS-Rosetta approach. In addition, a new CS-Rosetta protocol is described that improves robustness of the method for proteins with missing or erroneous NMR chemical shift input data. This strategy, which uses traditional Rosetta for pre-filtering of the fragment selection process, is demonstrated for two paramagnetic proteins and also for two proteins with solid-state NMR chemical shift assignments.