title: A temperature-jump NMR probe setup using rf heating optimized for the analysis of temperature-induced biomacromolecular kinetic processes.
abstract: A novel temperature jump (T-jump) probe operational at B(0) fields of 600 MHz (14.1 Tesla) with an integrated cage radio-frequency (rf) coil for rapid (<1 s) heating in high-resolution (HR) liquid-state NMR-spectroscopy is presented and its performance investigated. The probe consists of an inner 2.5 mm "heating coil" designed for generating rf-electric fields of 190-220 MHz across a lossy dielectric sample and an outer two coil assembly for (1)H-, (2)H- and (15)N-nuclei. High B(0) field homogeneities (0.7 Hz at 600 MHz) are combined with high heating rates (20-25 K/s) and only small temperature gradients (<±1.5 K, 3s after 20 K T-jump). The heating coil is under control of a high power rf-amplifier within the NMR console and can therefore easily be accessed by the pulse programmer. Furthermore, implementation of a real-time setup including synchronization of the NMR spectrometer's air flow heater with the rf-heater used to maintain the temperature of the sample is described. Finally, the applicability of the real-time T-jump setup for the investigation of biomolecular kinetic processes in the second-to-minute timescale is demonstrated for samples of a model 14mer DNA hairpin and a (15)N-selectively labeled 40nt hsp17-RNA thermometer.
authors.affiliation: Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe-University, Max-von-Laue-Strasse 7, D-60438 Frankfurt/Main, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Rinnenthal
authors.affiliation: Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe-University, Max-von-Laue-Strasse 7, D-60438 Frankfurt/Main, Germany.
authors.forename: Dominic
authors.initials: D
authors.lastname: Wagner
authors.affiliation: Bruker BioSpin GmbH, Am Silberstreifen, 76287 Rheinstetten, Germany.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Marquardsen
authors.affiliation: Bruker BioSpin GmbH, Am Silberstreifen, 76287 Rheinstetten, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Krahn
authors.affiliation: Bruker BioSpin GmbH, Am Silberstreifen, 76287 Rheinstetten, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Engelke
authors.affiliation: Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe-University, Max-von-Laue-Strasse 7, D-60438 Frankfurt/Main, Germany. Electronic address: schwalbe@nmr.uni-frankfurt.de.
authors.forename: Harald
authors.initials: H
authors.lastname: Schwalbe
mesh_terms: D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry; D004574:Electromagnetic Fields; D004581:Electronics; D006360:Heat-Shock Proteins / Q000737:chemistry; D007700:Kinetics; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000295:instrumentation*; D009690:Nucleic Acid Conformation; D011189:Potassium Chloride / Q000737:chemistry; D012313:RNA / Q000737:chemistry; D011846:Radio Waves; D013696:Temperature
chemical_list: D006360:Heat-Shock Proteins; C438147:heat-shock protein, 17-kDa; D012313:RNA; D011189:Potassium Chloride; D004247:DNA
keywords: Biomolecular unfolding; NMR spectroscopy; Radiofrequency heating; Temperature jump