Regulation of Eph receptor tyrosine kinase catalytic function.
Published: 2005
Language: eng
Number of pages: 187
Description: The eukaryotic protein kinases mediate diverse biological processes including metabolism, development, cell growth and differentiation. Accordingly, proper regulation of kinase catalytic activity is essential to the normal development and maintenance of eukaryotic organisms. A critical point of control for many protein kinases is the catalytic switch from an inactive state to an active state in response to specific stimuli. In the Eph receptor tyrosine kinase family, the catalytic switch is regulated by autophosphorylation within the juxtamembrane region (located between the transmembrane helix and the cytoplasmic kinase domain).I have solved the X-ray crystal structure of an autoinhibited, unphosphorylated form of EphB2 comprised of the juxtamembrane region and the kinase domain. As well, I have determined the crystal structure of the EphB2 kinase domain and the crystal structure of an active EphA4 mutant comprised of the juxtamembrane region and the kinase domain. The structures of Eph kinase domain in its autoinhibited and activated states, supported by mutagenesis data, reveal the molecular basis for Eph regulation by the juxtamembrane region. The unphosphorylated juxtamembrane region inhibits catalytic function by associating with the kinase domain, stabilizing the domain in a relatively open kinase conformation that compromises ATP coordination. In addition, the unphosphorylated juxtamembrane region sterically impedes the activation segment from attaining the productive conformation that is typical of active protein kinases, such that the juxtamembrane autoinhibitory structure and the activation segment are mutually exclusive. Phosphorylation of conserved juxtamembrane tyrosines would relieve this autoinhibition by disturbing the association of the juxtamembrane region with the kinase domain.My structural and mutational analyses of Eph kinase domains suggest that Tyr750 from the C-terminal kinase lobe also functions to control the conformation of the activation segment. It appears that the catalytic switch from the inactive state to the active state requires dissociation of the juxtamembrane region as well as repositioning of the side chain of Tyr750 to a conformation that is compatible with ordering of the activation segment. Interestingly, mutation of Tyr750 to alanine renders Eph activation independent of juxtamembrane autophosphorylation, suggesting that Tyr750Ala may behave as a gain-of-function mutation in Eph receptors.
Notes: Thesis (Ph.D.)--University of Toronto, 2005.

Electronic version licensed for access by U. of T. users.

Source: Dissertation Abstracts International, Volume: 66-06, Section: B, page: 2976.