PDBsum entry 2eb2

Transferase

PDB id: 2eb2

Contents

Protein chain

305 a.a. *

Waters ×16

* Residue conservation analysis

PDB id:

2eb2

Name:

Transferase

Title:

Crystal structure of mutated egfr kinase domain (g719s)

Structure:

Epidermal growth factor receptor. Chain: a. Fragment: kinase domain, unp residues 695-1022. Synonym: receptor tyrosine-protein kinase erbb-1. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: egfr. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108.

Resolution:

2.50Å R-factor: 0.198 R-free: 0.253

Authors:

S.Yoshikawa,M.Kukimoto-Niino,L.Chen,Z.J.Liu,B.C.Wang,M.Shirouzu, K.Senba,T.Yamamoto,S.Yokoyama,Riken Structural Genomics/proteomics Initiative (Rsgi)

Key ref:

S.Yoshikawa et al. (2013). Structural basis for the altered drug sensitivities of non-small cell lung cancer-associated mutants of human epidermal growth factor receptor. Oncogene, 32, 27-38. PubMed id: 22349823

Date:

06-Feb-07 Release date: 12-Feb-08

Protein chain

P00533  (EGFR_HUMAN) -  Epidermal growth factor receptor from Homo sapiens

Seq: 1210 a.a.

Struc: 305 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.7.10.1 - receptor 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

Oncogene 32:27-38 (2013)

PubMed id: 22349823

Structural basis for the altered drug sensitivities of non-small cell lung cancer-associated mutants of human epidermal growth factor receptor.

S.Yoshikawa, M.Kukimoto-Niino, L.Parker, N.Handa, T.Terada, T.Fujimoto, Y.Terazawa, M.Wakiyama, M.Sato, S.Sano, T.Kobayashi, T.Tanaka, L.Chen, Z.J.Liu, B.C.Wang, M.Shirouzu, S.Kawa, K.Semba, T.Yamamoto, S.Yokoyama.

ABSTRACT

The epidermal growth factor receptor (EGFR) has an essential role in multiple signaling pathways, including cell proliferation and migration, through extracellular ligand binding and subsequent activation of its intracellular tyrosine kinase (TK) domain. The non-small cell lung cancer (NSCLC)-associated EGFR mutants, L858R and G719S, are constitutively active and oncogenic. They display sensitivity to TK inhibitors, including gefitinib and erlotinib. In contrast, the secondary mutation of the gatekeeper residue, T790M, reportedly confers inhibitor resistance on the oncogenic EGFR mutants. In this study, our biochemical analyses revealed that the introduction of the T790M mutation confers gefitinib resistance on the G719S mutant. The G719S/T790M double mutant has enhanced activity and retains high gefitinib-binding affinity. The T790M mutation increases the ATP affinity of the G719S mutant, explaining the acquired drug resistance of the double mutant. Structural analyses of the G719S/T790M double mutant, as well as the wild type and the G719S and L858R mutants, revealed that the T790M mutation stabilizes the hydrophobic spine of the active EGFR-TK conformation. The Met790 side chain of the G719S/T790M double mutant, in the apo form and gefitinib- and AMPPNP-bound forms, adopts different conformations that explain the accommodation of these ligands. In the L858R mutant structure, the active-site cleft is expanded by the repositioning of Phe723 within the P-loop. Notably, the introduction of the F723A mutation greatly enhanced the gefitinib sensitivity of the wild-type EGFR in vivo, supporting our hypothesis that the expansion of the active-site cleft results in enhanced gefitinib sensitivity. Taken together, our results provide a structural basis for the altered drug sensitivities caused by distinct NSCLC-associated EGFR mutations.