Structural changes in the Ras protein revealed by fluorescence spectroscopy

Brockhinke A, Plessow R, Kohse-Höinghaus K, Herrmann C (2003)
PHYSICAL CHEMISTRY CHEMICAL PHYSICS 5(16): 3498-3506.

Journal Article | Published | English

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Abstract
The Y32W mutant of the Ras protein which has a tryptophan residue close to the guanine nucleotide binding site is studied using two fluorescence spectroscopic techniques. Two-dimensional mapping of all emission and all fluorescence spectra using excitation-emission spectroscopy (EES) in conjunction with time-resolved laser-induced fluorescence (LIF) is used to analyze and assign the contribution of the different fluorophores to the total fluorescence. Time-resolved LIF is shown to be a method that allows to follow the slight conformational changes of Ras binding to the nucleotides GDP, GTP, or the non-hydrolyzable analogues GppNHp, GppCH(2)p and GTP-gammaS and allows to distinguish between the active and inactive form. Additionally, a variant of the EES technique is used for the investigation of the intrinsic GTPase function of Ras and the determination of kinetic constants for this reaction.
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Brockhinke A, Plessow R, Kohse-Höinghaus K, Herrmann C. Structural changes in the Ras protein revealed by fluorescence spectroscopy. PHYSICAL CHEMISTRY CHEMICAL PHYSICS. 2003;5(16):3498-3506.
Brockhinke, A., Plessow, R., Kohse-Höinghaus, K., & Herrmann, C. (2003). Structural changes in the Ras protein revealed by fluorescence spectroscopy. PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 5(16), 3498-3506.
Brockhinke, A., Plessow, R., Kohse-Höinghaus, K., and Herrmann, C. (2003). Structural changes in the Ras protein revealed by fluorescence spectroscopy. PHYSICAL CHEMISTRY CHEMICAL PHYSICS 5, 3498-3506.
Brockhinke, A., et al., 2003. Structural changes in the Ras protein revealed by fluorescence spectroscopy. PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 5(16), p 3498-3506.
A. Brockhinke, et al., “Structural changes in the Ras protein revealed by fluorescence spectroscopy”, PHYSICAL CHEMISTRY CHEMICAL PHYSICS, vol. 5, 2003, pp. 3498-3506.
Brockhinke, A., Plessow, R., Kohse-Höinghaus, K., Herrmann, C.: Structural changes in the Ras protein revealed by fluorescence spectroscopy. PHYSICAL CHEMISTRY CHEMICAL PHYSICS. 5, 3498-3506 (2003).
Brockhinke, Andreas, Plessow, Regina, Kohse-Höinghaus, Katharina, and Herrmann, Christian. “Structural changes in the Ras protein revealed by fluorescence spectroscopy”. PHYSICAL CHEMISTRY CHEMICAL PHYSICS 5.16 (2003): 3498-3506.
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