Amide additives improve RDC measurements in polyacrylamide
| Autor: | Jun Zhang, Zihan Zhang, Taylor Knapp, Naiduwadura Ivon Upekala De Silva, Heather McKelvey, Talia Fargason, Steve Zaharias, Ting Wang |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Chemistry Polyacrylamide Acrylic Resins 010402 general chemistry 01 natural sciences Biochemistry 0104 chemical sciences 03 medical and health sciences chemistry.chemical_compound Crystallography 030104 developmental biology Protein stability Amide Chemical stability Asparagine Protein solubility Nuclear Magnetic Resonance Biomolecular Polyacrylamide gel electrophoresis Spectroscopy Macromolecule |
| Zdroj: | Journal of Biomolecular NMR. 74:119-124 |
| ISSN: | 1573-5001 0925-2738 |
| Popis: | Residual dipolar couplings (RDCs) provide valuable NMR parameters that can be used for structural calculation and verification. Measuring RDCs requires aligning macromolecules using one of various types of alignment media. Of different alignment media options, stretched or compressed polyacrylamide gels are advantageous due to their chemical stability. However, polyacrylamide interacts with proteins and significantly broadens NMR resonances. In this study, we found that the amide-containing compounds asparagine, glutamine and propionamide improve spectral quality of proteins in polyacrylamide gel without significantly reducing the magnitude of RDC values. Moreover, we showed that propionamide is an attractive additive that increases protein solubility without interfering with protein stability, ligand binding or NMR pulse width, suggesting its potential applications for our NMR methods. |
| Databáze: | OpenAIRE |
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