Structural basis for the recognition of nucleophosmin-anaplastic lymphoma kinase oncoprotein by the phosphotyrosine binding domain of Suc1-associated neurotrophic factor-induced tyrosine-phosphorylated target-2

• Published in: Journal of structural and functional genomics Vol. 11; no. 2; pp. 125 - 141
• Main Authors: Koshiba, Seizo, Li, Hua, Motoda, Yoko, Tomizawa, Tadashi, Kasai, Takuma, Tochio, Naoya, Yabuki, Takashi, Harada, Takushi, Watanabe, Satoru, Tanaka, Akiko, Shirouzu, Mikako, Kigawa, Takanori, Yamamoto, Tadashi, Yokoyama, Shigeyuki
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01.06.2010; Springer Netherlands; Springer Nature B.V
• Subjects: Adaptor Proteins, Signal Transducing - metabolism; Amino Acid Sequence; Amino acids; Binding Sites; Biochemistry; Bioinformatics; Biomedical and Life Sciences; Crystallography, X-Ray; Fibroblasts; Forestry Management; Growth factors; Human Genetics; Humans; Kinases; Life Sciences; Lymphoma; Magnetic Resonance Spectroscopy; Microbial Genetics and Genomics; Molecular Sequence Data; NMR; NPM-ALK; Nuclear magnetic resonance; Peptide Fragments; Peptides; Phosphorylation; Phosphotyrosine - metabolism; Plant Genetics and Genomics; Protein Binding; Protein Conformation; Protein expression; Protein Structure, Tertiary; Protein-Tyrosine Kinases - chemistry; Protein-Tyrosine Kinases - genetics; Protein-Tyrosine Kinases - metabolism; Proteins; PTB domain; Science; Sequence Homology, Amino Acid; SNT-2; Spectrum analysis; Tyrosine kinase; Vectors (Biology); SNT-2; Tyrosine kinase; NMR; NPM-ALK; PTB domain
• ISSN: 1345-711X; 1570-0267
• DOI: 10.1007/s10969-010-9091-x

The nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) fusion oncoprotein, formed by the t(2;5) chromosomal translocation in anaplastic large-cell lymphomas, has constitutive tyrosine kinase activity and interacts with a number of signaling molecules. One of the interacting partners of NPM-ALK is the adaptor protein, Suc1-associated neurotrophic factor-induced tyrosine-phosphorylated target (SNT), and mutations that deprive NPM-ALK of all three of the SNT-binding sites significantly reduced the transforming activity. In this study, the interactions of the three binding sites in NPM-ALK with the phosphotyrosine binding (PTB) domain of SNT-2 were analyzed. First, by isothermal titration calorimetry, we found that the phosphorylation-independent binding site in NPM-ALK interacts with the SNT-2 PTB domain more tightly than the phosphorylation-dependent binding sites. Second, the solution structure of the SNT-2 PTB domain in complex with the nonphosphorylated NPM-ALK peptide was determined by nuclear magnetic resonance spectroscopy. The NPM-ALK peptide interacts with the hydrophobic surface of the PTB domain and intermolecularly extends the PTB β-sheet. This interaction mode is much broader and more extensive than those of the phosphorylation-dependent binding sites. Our results indicate that the higher binding activity of the phosphorylation-independent binding site is caused by additional hydrophobic interactions.