5Y8U

Crystal structure of the C276S mutant of MAP2K7

Summary for 5Y8U

• Entry DOI: 10.2210/pdb5y8u/pdb
• Descriptor: Dual specificity mitogen-activated protein kinase kinase 7 (2 entities in total)
• Functional Keywords: transferase, protein kinase
• Biological source: Homo sapiens (Human)
• Cellular location: Nucleus: O14733
• Total number of polymer chains: 1
• Total formula weight: 36851.64

Authors

Kinoshita, T. (deposition date: 2017-08-21, release date: 2017-10-11, Last modification date: 2023-11-22)

Primary citation

Kinoshita, T.,Hashimoto, T.,Sogabe, Y.,Fukada, H.,Matsumoto, T.,Sawa, M.
High-resolution structure discloses the potential for allosteric regulation of mitogen-activated protein kinase kinase 7
Biochem. Biophys. Res. Commun., 493:313-317, 2017

PubMed Abstract: Mitogen-activated protein kinase kinase 7 (MAP2K7) regulates stress and inflammatory responses, and is an attractive drug discovery target for several diseases including arthritis and cardiac hypertrophy. Intracellular proteins such as MAP2K7 are prone to aggregation due to cysteine-driven oxidation in in vitro experiments. MAP2K7 instability due to the four free cysteine residues on the molecular surface abrogated the crystal growth and led to a low-resolution structure with large residual errors. To acquire a higher resolution structure for promoting rational drug discovery, we explored stable mutants of MAP2K7 by replacing the surface cysteine residues, Cys147, Cys218, Cys276 and Cys296. Single-site mutations, except for Cys147, maintained the specific activity and increased the protein yield, while all the multi-site mutations massively reduced the activity. The C218S mutation drastically augmented the protein production and crystallographic resolution. Furthermore, the C218S crystals grown under microgravity in a space environment yielded a 1.3 Å resolution structure, providing novel insights for drug discovery: the precisely assigned water molecules in the active site, the double conformations in the flexible region and the C-terminal extension bound to the N-terminal region of the adjacent molecules. The latter insight is likely to promote the production of allosteric MAP2K7 inhibitors.

PubMed: 28890347
DOI: 10.1016/j.bbrc.2017.09.025

Experimental method

X-RAY DIFFRACTION (2.92 Å)