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演 題 「Mechanistic studies of the Ras-superfamily by FTIR spectroscopy」
日 時 2013年08月30日(金) 16:00 より 18:00 まで
講演者 PD Dr. Carsten Kötting Permanent Position "PD", Department of Biology and Biotechnology, Ruhr-University Bochum
場 所

Yamate Building 3, 2nd floor, Seminar Room

概 要

The Ras protein is mutated in 30% of human tumors. Ras acts as a switch, transmitting a growth signal in an active GTP-bound form and turning the signal off in an inactive GDP-bound form. The switch-off is accomplished by GTP hydrolysis, which is catalyzed by Ras and can be further accelerated by GTPase activating proteins (GAPs). Mutations which inhibit hydrolysis cause severe diseases including cancer. We investigated the reaction of the Ras·GAP protein-protein complex by time-resolved FTIR spectroscopy by means of caged-GTP.1 This method was also applied to other GTPases.2
Further, we optimized an ATR-FTIR system based on a germanium crystal to perform stimulus induced IR difference spectroscopy of monolayers without the surface enhance effect. We synthesized linker molecules which allow for stable attachment of proteins via tags.3 In the flow thorough system protein-ligand, protein-protein and protein-drug interactions can be investigated. Alternatively a model membrane can be attached to the germanium and membrane anchored proteins such as the GTPases Ras and Rab can be investigated in its natural environment. We found that N-Ras dimerizes at a POPC model membrane.4 For Rab, the membrane extraction by GDI was investigated.5

1Kötting, Güldenhaupt, Gerwert, Chemical Physics 396, 72 (2012). 2Kötting, Gerwert, FEBS Letters 587, 2025 (2013) 3Schartner et al., JACS 135, 4079 (2013). 4Güldenhaupt et al., Biophys. J. 103, 1585 (2012). 5Gavriljuk et al., PNAS, in press.

お問合せ先

Yuji Furutani