PDBsum entry 4e1z

Transferase/transferase inhibitor

PDB id: 4e1z

Contents

Protein chain

288 a.a.

Ligands

0MX

Waters ×41

PDB id:

4e1z

Name:

Transferase/transferase inhibitor

Title:

Structure of mouse tyk-2 complexed to a 3-aminoindazole inhibitor

Structure:

Non-receptor tyrosine-protein kinase tyk2. Chain: a. Fragment: tyk-2 catalytic domain (unp residues 880-1170). Engineered: yes. Mutation: yes

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Gene: tyk2. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108

Resolution:

2.50Å R-factor: 0.199 R-free: 0.237

Authors:

M.A.Argiriadi,R.V.Talanian,D.W.Borhani

Key ref:

M.A.Argiriadi et al. (2012). Enabling structure-based drug design of Tyk2 through co-crystallization with a stabilizing aminoindazole inhibitor. Bmc Struct Biol, 12, 22. PubMed id: 22995073

Date:

07-Mar-12 Release date: 03-Oct-12

Protein chain

Q9R117  (TYK2_MOUSE) -  Non-receptor tyrosine-protein kinase TYK2 from Mus musculus

Seq: 1184 a.a.

Struc: 288 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.7.10.2 - non-specific protein-tyrosine kinase.
• Reaction: L-tyrosyl-[protein] + ATP = O-phospho-L-tyrosyl-[protein] + ADP + H⁺

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

Bmc Struct Biol 12:22 (2012)

PubMed id: 22995073

Enabling structure-based drug design of Tyk2 through co-crystallization with a stabilizing aminoindazole inhibitor.

M.A.Argiriadi, E.R.Goedken, D.Banach, D.W.Borhani, A.Burchat, R.W.Dixon, D.Marcotte, G.Overmeyer, V.Pivorunas, R.Sadhukhan, S.Sousa, N.S.Moore, M.Tomlinson, J.Voss, L.Wang, N.Wishart, K.Woller, R.V.Talanian.

ABSTRACT

ABSTRACT: BACKGROUND: Structure-based drug design (SBDD) can accelerate inhibitor lead design and optimization, and efficient methods including protein purification, characterization, crystallization, and high-resolution diffraction are all needed for rapid, iterative structure determination. Janus kinases are important targets that are amenable to structure-based drug design. Here we present the first mouse Tyk2 crystal structures, which are complexed to 3-aminoindazole compounds. RESULTS: A comprehensive construct design effort included N- and C-terminal variations, kinase-inactive mutations, and multiple species orthologs. High-throughput cloning and expression methods were coupled with an abbreviated purification protocol to optimize protein solubility and stability. In total, 50 Tyk2 constructs were generated. Many displayed poor expression, inadequate solubility, or incomplete affinity tag processing. One kinase-inactive murine Tyk2 construct, complexed with an ATP-competitive 3-aminoindazole inhibitor, provided crystals that diffracted to 2.5--2.6 A resolution. This structure revealed initial "hot-spot" regions for SBDD, and provided a robust platform for ligand soaking experiments. Compared to previously reported human Tyk2 inhibitor crystal structures (Chrencik et al. (2010) J Mol Biol 400:413), our structures revealed a key difference in the glycine-rich loop conformation that is induced by the inhibitor. Ligand binding also conferred resistance to proteolytic degradation by thermolysin. As crystals could not be obtained with the unliganded enzyme, this enhanced stability is likely important for successful crystallization and inhibitor soaking methods. CONCLUSIONS: Practical criteria for construct performance and prioritization, the optimization of purification protocols to enhance protein yields and stability, and use of high-throughput construct exploration enable structure determination methods early in the drug discovery process. Additionally, specific ligands stabilize Tyk2 protein and may thereby enable crystallization.