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.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
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.
Erscheinungsjahr
Zeitschriftentitel
PHYSICAL CHEMISTRY CHEMICAL PHYSICS
Band
5
Zeitschriftennummer
16
Seite
3498-3506
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eISSN
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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. doi:10.1039/b303262k
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.