title: Binding, domain orientation, and dynamics of the Lck SH3-SH2 domain pair and comparison with other Src-family kinases.
abstract: The catalytic activity of Src-family kinases is regulated by association with its SH3 and SH2 domains. Activation requires displacement of intermolecular contacts by SH3/SH2 binding ligands resulting in dissociation of the SH3 and SH2 domains from the kinase domain. To understand the contribution of the SH3-SH2 domain pair to this regulatory process, the binding of peptides derived from physiologically relevant SH2 and SH3 interaction partners was studied for Lck and its relative Fyn by NMR spectroscopy. In contrast to Fyn, activating ligands do not induce communication between SH2 and SH3 domains in Lck. This can be attributed to the particular properties of the Lck SH3-SH2 linker which is shown to be extremely flexible thus effectively decoupling the behavior of the SH3 and SH2 domains. Measurements on the SH32 tandem from Lck further revealed a relative domain orientation that is distinctly different from that found in the Lck SH32 crystal structure and in other Src kinases. These data suggest that flexibility between SH2 and SH3 domains contributes to the adaptation of Src-family kinases to specific environments and distinct functions.
authors.affiliation: Department of Biochemistry, University of Oxford, UK.
authors.forename: Gregor
authors.initials: G
authors.lastname: Hofmann
authors.forename: Kristian
authors.initials: K
authors.lastname: Schweimer
authors.forename: Anke
authors.initials: A
authors.lastname: Kiessling
authors.forename: Edith
authors.initials: E
authors.lastname: Hofinger
authors.forename: Finn
authors.initials: F
authors.lastname: Bauer
authors.forename: Silke
authors.initials: S
authors.lastname: Hoffmann
authors.forename: Paul
authors.initials: P
authors.lastname: Rösch
authors.forename: Iain D
authors.initials: ID
authors.lastname: Campbell
authors.forename: Jörn M
authors.initials: JM
authors.lastname: Werner
authors.forename: Heinrich
authors.initials: H
authors.lastname: Sticht
mesh_terms: D006801:Humans; D008024:Ligands; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000737:chemistry* / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051076:Proto-Oncogene Proteins c-fyn / Q000737:chemistry; D018909:src Homology Domains; D019061:src-Family Kinases / Q000737:chemistry*
chemical_list: D008024:Ligands; D010455:Peptides; C494957:FYN protein, human; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051076:Proto-Oncogene Proteins c-fyn; D019061:src-Family Kinases