6QTY
Non-phosphorylated human CLK1 in complex with an indole inhibitor to 1.65 Ang
Summary for 6QTY
| Entry DOI | 10.2210/pdb6qty/pdb |
| Related | 2VAG |
| Descriptor | Dual specificity protein kinase CLK1, ethyl 3-[(E)-2-amino-1-cyanoethenyl]-6,7-dichloro-1-methyl-1H-indole-2-carboxylate (3 entities in total) |
| Functional Keywords | clk1, splicing, kinase, viral infection, signaling protein |
| Biological source | Homo sapiens (Human) |
| Total number of polymer chains | 1 |
| Total formula weight | 39787.54 |
| Authors | Livnah, O.,Domovich, Y. (deposition date: 2019-02-26, release date: 2020-03-18, Last modification date: 2024-01-24) |
| Primary citation | Dekel, N.,Eisenberg-Domovich, Y.,Karlas, A.,Meyer, T.F.,Bracher, F.,Lebendiker, M.,Danieli, T.,Livnah, O. Expression, purification and crystallization of CLK1 kinase - A potential target for antiviral therapy. Protein Expr.Purif., 176:105742-105742, 2020 Cited by PubMed Abstract: Cdc-like kinase 1 (CLK1) is a dual-specificity kinase capable of autophosphorylation on tyrosine residues and Ser/Thr phosphorylation of its substrates. CLK1 belongs to the CLK kinase family that regulates alternative splicing through phosphorylation of serine-arginine rich (SR) proteins. Recent studies have demonstrated that CLK1 has an important role in the replication of influenza A and chikungunya viruses. Furthermore, CLK1 was found to be relevant for the replication of HIV-1 and the West Nile virus, making CLK1 an interesting cellular candidate for the development of a host-directed antiviral therapy that might be efficient for treatment of newly emerging viruses. We describe here our attempts and detailed procedures to obtain the recombinant kinase domain of CLK1 in suitable amounts for crystallization in complex with specific inhibitors. The key solution for the reproducibility of crystals resides in devising and refining expression and purification protocols leading to homogeneous protein. Co-expression of CLK1 with λ-phosphatase and careful purification has yielded crystals of CLK1 complexed with the KH-CB19 inhibitor that diffracted to 1.65 Å. These results paved the path to the screening of more structures of CLK1 complexed compounds, leading to further optimization of their inhibitory activity. Moreover, since kinases are desired targets in numerous pathologies, the approach we report here, the co-expression of kinases with λ-phosphatase, previously used in other kinases, can be adopted as a general protocol in numerous kinase targets for obtaining reproducible and homogenic non-phosphorylated (inactive) forms suitable for biochemical and structural studies thus facilitating the development of novel inhibitors. PubMed: 32866611DOI: 10.1016/j.pep.2020.105742 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.65 Å) |
Structure validation
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