4TT7

Crystal structure of human ALK with a covalent modification

Summary for 4TT7

• Entry DOI: 10.2210/pdb4tt7/pdb
• Descriptor: ALK tyrosine kinase receptor, 1-THIOETHANESULFONIC ACID, 1,2-ETHANEDIOL, ... (4 entities in total)
• Functional Keywords: atp-binding, receptor, transferase
• Biological source: Homo sapiens (Human)
• Cellular location: Cell membrane ; Single-pass type I membrane protein : Q9UM73
• Total number of polymer chains: 1
• Total formula weight: 37240.79

Authors

Badger, J.,Sridhar, V.,Chie-Leon, B.,Nienaber, V.L.,Hausheer, F.H. (deposition date: 2014-06-19, release date: 2015-02-11, Last modification date: 2024-10-23)

Primary citation

Parker, A.R.,Petluru, P.N.,Nienaber, V.L.,Zhao, M.,Ayala, P.Y.,Badger, J.,Chie-Leon, B.,Sridhar, V.,Logan, C.,Kochat, H.,Hausheer, F.H.
Novel covalent modification of human anaplastic lymphoma kinase (ALK) and potentiation of crizotinib-mediated inhibition of ALK activity by BNP7787.
Onco Targets Ther, 8:375-383, 2015

PubMed Abstract: BNP7787 (Tavocept, disodium 2,2'-dithio-bis-ethanesulfonate) is a novel, investigational, water-soluble disulfide that is well-tolerated and nontoxic. In separate randomized multicenter Phase II and Phase III clinical trials in non-small-cell lung cancer (NSCLC) patients, treatment with BNP7787 in combination with standard chemotherapy resulted in substantial increases in the overall survival of patients with advanced adenocarcinoma of the lung in the first-line treatment setting. We hypothesized that BNP7787 might interact with and modify human anaplastic lymphoma kinase (ALK). At least seven different variants of ALK fusions with the gene encoding the echinoderm microtubule-associated protein-like 4 (EML4) are known to occur in NSCLC. EML4-ALK fusions are thought to account for approximately 3% of NSCLC cases. Herein, we report the covalent modification of the kinase domain of human ALK by a BNP7787-derived mesna moiety and the functional consequences of this modification in ALK assays evaluating kinase activity. The kinase domain of the ALK protein crystallizes as a monomer, and BNP7787-derived mesna-cysteine adducts were observed at Cys 1235 and Cys 1156. The BNP7787-derived mesna adduct at Cys 1156 is located in close proximity to the active site and results in substantial disorder of the P-loop and activation loop (A-loop). Comparison with the P-loop of apo-ALK suggests that the BNP7787-derived mesna adduct at Cys 1156 interferes with the positioning of Phe 1127 into a small pocket now occupied by mesna, resulting in a destabilization of the loop's binding orientation. Additionally, in vitro kinase activity assays indicate that BNP7787 inhibits ALK catalytic activity and potentiates the activity of the ALK-targeted drug crizotinib.

PubMed: 25678804
DOI: 10.2147/OTT.S73690

Experimental method

X-RAY DIFFRACTION (2.1 Å)