Recombinant Expression and Purification of the RNA-binding LARP6 Proteins from Fish Genetic Model Organisms
| dc.contributor.author | Castro, Jose M. | |
| dc.contributor.author | Horn, Daniel | |
| dc.contributor.author | Lewis, Karen A. | |
| dc.date.accessioned | 2020-04-02T16:54:50Z | |
| dc.date.available | 2020-04-02T16:54:50Z | |
| dc.date.issued | 2017-06 | |
| dc.description.abstract | The RNA-binding proteins that comprise the La-related protein (LARP) superfamily have been implicated in a wide range of cellular functions, from tRNA maturation to regulation of protein synthesis. To more expansively characterize the biological function of the LARP6 subfamily, we have recombinantly expressed the full-length LARP6 proteins from two teleost fish, platyfish (Xiphophorus maculatus) and zebrafish (Danio rerio). The yields of the recombinant proteins were enhanced to >2 mg/L using a tandem approach of an N-terminal His6-SUMO tag and an iterative solubility screening assay to identify structurally stabilizing buffer components. The domain topologies of the purified fish proteins were probed with limited proteolysis. The fish proteins contain an internal, protease-resistant 40 kDa domain, which is considerably more stable than the comparable domain from the human LARP6 protein. The fish proteins are therefore a lucrative model system in which to study both the evolutionary divergence of this family of La-related proteins and the structure and conformational dynamics of the domains that comprise the LARP6 protein. | |
| dc.description.department | Chemistry and Biochemistry | |
| dc.description.version | This is the accepted manuscript version of an article published in Protein Expression and Purification. | |
| dc.format | Text | |
| dc.format.extent | 14 pages | |
| dc.format.medium | 1 file (.pdf) | |
| dc.identifier.citation | Castro, J. M., Horn, D. A., & Lewis, K. A. (2017). Recombinant expression and purification of the RNA-binding LARP6 proteins from fish genetic model organisms. Protein Expression and Purification, 134, pp. 147–153. | |
| dc.identifier.doi | https://doi.org/10.1016/j.pep.2017.04.004 | |
| dc.identifier.issn | 1046-5928 | |
| dc.identifier.uri | https://hdl.handle.net/10877/9572 | |
| dc.language.iso | en | |
| dc.publisher | Elsevier | |
| dc.source | Protein Expression and Purification, 2017, Vol. 134, pp. 147–153. | |
| dc.subject | la-related protein | |
| dc.subject | bacterial expression | |
| dc.subject | protein solubility | |
| dc.subject | RNA binding protein | |
| dc.subject | Chemistry and Biochemistry | |
| dc.title | Recombinant Expression and Purification of the RNA-binding LARP6 Proteins from Fish Genetic Model Organisms | |
| dc.type | Article |