title: Identification of a new interaction mode between the Src homology 2 domain of C-terminal Src kinase (Csk) and Csk-binding protein/phosphoprotein associated with glycosphingolipid microdomains.
abstract: Proteins with Src homology 2 (SH2) domains play major roles in tyrosine kinase signaling. Structures of many SH2 domains have been studied, and the regions involved in their interactions with ligands have been elucidated. However, these analyses have been performed using short peptides consisting of phosphotyrosine followed by a few amino acids, which are described as the canonical recognition sites. Here, we report the solution structure of the SH2 domain of C-terminal Src kinase (Csk) in complex with a longer phosphopeptide from the Csk-binding protein (Cbp). This structure, together with biochemical experiments, revealed the existence of a novel binding region in addition to the canonical phosphotyrosine 314-binding site of Cbp. Mutational analysis of this second region in cells showed that both canonical and novel binding sites are required for tumor suppression through the Cbp-Csk interaction. Furthermore, the data indicate an allosteric connection between Cbp binding and Csk activation that arises from residues in the βB/βC loop of the SH2 domain.
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Akagi
authors.forename: Chitose
authors.initials: C
authors.lastname: Oneyama
authors.forename: Masakazu
authors.initials: M
authors.lastname: Tanaka
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Sasaki
authors.forename: Takashi
authors.initials: T
authors.lastname: Kanou
authors.forename: Young-Ho
authors.initials: YH
authors.lastname: Lee
authors.forename: Daisuke
authors.initials: D
authors.lastname: Yokogawa
authors.forename: Marc-Werner
authors.initials: MW
authors.lastname: Dobenecker
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nakagawa
authors.forename: Masato
authors.initials: M
authors.lastname: Okada
authors.forename: Takahisa
authors.initials: T
authors.lastname: Ikegami
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D000081247:CSK Tyrosine-Protein Kinase; D006028:Glycosphingolipids / Q000235:genetics / Q000378:metabolism*; D021962:Membrane Microdomains / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D051381:Rats; D061987:Sf9 Cells; D018411:Spodoptera; D018909:src Homology Domains; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D006028:Glycosphingolipids; D010750:Phosphoproteins; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases
keywords: C-terminal Src Kinase; Csk-binding protein; NMR; Protein Structure; SH2 Domains; Src; Tumor; Tyrosine Protein Kinase (Tyrosine Kinase)
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