A Method for Systematic Assessment of Intrinsically Disordered Protein Regions by NMR
AbstractIntrinsically disordered proteins (IDPs) that lack stable conformations and are highly flexible have attracted the attention of biologists. Therefore, the development of a systematic method to identify polypeptide regions that are unstructured in solution is important. We have designed an “indirect/reflected” detection system for evaluating the physicochemical properties of IDPs using nuclear magnetic resonance (NMR). This approach employs a “chimeric membrane protein”-based method using the thermostable membrane protein PH0471. This protein contains two domains, a transmembrane helical region and a C-terminal OB (oligonucleotide/oligosaccharide binding)-fold domain (named NfeDC domain), connected by a flexible linker. NMR signals of the OB-fold domain of detergent-solubilized PH0471 are observed because of the flexibility of the linker region. In this study, the linker region was substituted with target IDPs. Fifty-three candidates were selected using the prediction tool POODLE and 35 expression vectors were constructed. Subsequently, we obtained 15N-labeled chimeric PH0471 proteins with 25 IDPs as linkers. The NMR spectra allowed us to classify IDPs into three categories: flexible, moderately flexible, and inflexible. The inflexible IDPs contain membrane-associating or aggregation-prone sequences. This is the first attempt to use an indirect/reflected NMR method to evaluate IDPs and can verify the predictions derived from our computational tools. View Full-Text
- Supplementary File 1:
Supplementary (PDF, 752 KB)
Share & Cite This Article
Goda, N.; Shimizu, K.; Kuwahara, Y.; Tenno, T.; Noguchi, T.; Ikegami, T.; Ota, M.; Hiroaki, H. A Method for Systematic Assessment of Intrinsically Disordered Protein Regions by NMR. Int. J. Mol. Sci. 2015, 16, 15743-15760.
Goda N, Shimizu K, Kuwahara Y, Tenno T, Noguchi T, Ikegami T, Ota M, Hiroaki H. A Method for Systematic Assessment of Intrinsically Disordered Protein Regions by NMR. International Journal of Molecular Sciences. 2015; 16(7):15743-15760.Chicago/Turabian Style
Goda, Natsuko; Shimizu, Kana; Kuwahara, Yohta; Tenno, Takeshi; Noguchi, Tamotsu; Ikegami, Takahisa; Ota, Motonori; Hiroaki, Hidekazu. 2015. "A Method for Systematic Assessment of Intrinsically Disordered Protein Regions by NMR." Int. J. Mol. Sci. 16, no. 7: 15743-15760.