Gao, YunrongCao, DongdongPawnikar, ShristiAkhter, SanaMiao, YinglongLiang, Bo2022-01-072022-01-072021-03-19Gao, Y., Cao, D., Pawnikar, S., Akhter, S., Miao, Y., & Liang, B. (2021). Efficient purification and assembly of ribonucleoprotein complex for interaction analysis by MST assay coupled with GaMD simulations. STAR protocols, 2(1), 100315. https://doi.org/10.1016/j.xpro.2021.100315https://hdl.handle.net/1808/32371Here, we describe a generic protocol for monitoring protein-RNA interaction using a cleavable GFP fusion of a recombinant RNA-binding protein. We detail each expression and purification step, including high salt and heparin column for contaminant RNA removal. After the assembly of RNA into the ribonucleoprotein complex, the MicroScale Thermophoresis assay enables the binding affinity to be obtained quickly with a small amount of sample. Further Gaussian accelerated molecular dynamics simulations allow us to analyze protein:RNA interactions in detail.Copyright: 2021 The Author(s). This is an open access article under the CC BY-NC-ND license.http://creativecommons.org/licenses/by-nc-nd/4.0/BioinformaticsProtein biochemistryProtein expression and purificationEfficient purification and assembly of ribonucleoprotein complex for interaction analysis by MST assay coupled with GaMD simulationsArticle10.1016/j.xpro.2021.100315PMC7890042openAccess