Time-resolved flow-flash FT-IR difference spectroscopy: the kinetics of CO photodissociation from myoglobin revisited

Schleeger, Michael; Wagner, Christoph; Vellekoop, Michael J.; Lendl, Bernhard; Heberle, Joachim

doi:10.1007/s00216-009-2928-0

Cited by 4 publications

(6 citation statements)

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“…We have developed a spectrometer based on tunable QCLs for recording time-resolved difference spectra in the mid-IR range. The recorded data from the model systems bR and MbCO agree well with published time-resolved FT-IR spectroscopy experiments using the step-scan technique [29,30,53]. The major advantage of our QCL setup is the straightforward measurement of strong absorbing samples which relates to the high emission power of QCLs.…”

Section: Discussionsupporting

confidence: 79%

“…12b. The latter data was taken within 25 seconds which compares to more than 1 hour for a step scan FT-IR experiment recorded under otherwise identical conditions [30]. However, this comparison is only valid if spectral information on particular marker bands has already been retrived.…”

Section: Mbco In Aqueous Solutionmentioning

confidence: 99%

“…In particular, bR has been used in pioneering time-resolved IR studies ranging from fs pumpprobe experiments [28] to step-scan FTIR with microsecond and nanosecond resolution [29]. As a model metalloprotein, myoglobin (Mb) was employed studying the photo-dissociation of bound carbonmonoxide (CO) [30][31][32].…”

Section: Introductionmentioning

confidence: 99%

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Protein dynamics observed by tunable mid-IR quantum cascade lasers across the time range from 10 ns to 1 s

Schultz

Mohrmann

Lórenz-Fonfrı́a

et al. 2018

Spectrochimica Acta Part A: Molecular and Biomolecular Spectros

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We have developed a spectrometer based on tunable quantum cascade lasers (QCLs) for recording time-resolved absorption spectra of proteins in the mid-infrared range. We illustrate its performance by recording time-resolved difference spectra of bacteriorhodopsin in the carboxylic range (1800-1700cm) and on the CO rebinding reaction of myoglobin (1960-1840cm), at a spectral resolution of 1cm. The spectrometric setup covers the time range from 4ns to nearly a second with a response time of 10-15ns. Absorption changes as low as 1×10 are detected in single-shot experiments at t>1μs, and of 5×10 in kinetics obtained after averaging 100 shots. While previous time-resolved IR experiments have mostly been conducted on hydrated films of proteins, we demonstrate here that the brilliance of tunable quantum cascade lasers is superior to perform ns time-resolved experiments even in aqueous solution (HO).

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Section: Discussionsupporting

confidence: 79%

Section: Mbco In Aqueous Solutionmentioning

confidence: 99%

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Protein dynamics observed by tunable mid-IR quantum cascade lasers across the time range from 10 ns to 1 s

Schultz

Mohrmann

Lórenz-Fonfrı́a

et al. 2018

Spectrochimica Acta Part A: Molecular and Biomolecular Spectros

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“…Therefore, probing the dynamic structural information during a non-equilibrium process holds a great promise to understand the mechanism of biological reactions driven by variation of the chemical/physical environment of a biological system. Very recently, pioneering groups attempted to develop time-resolved techniques to monitor the catalytic mechanism of proteins during reactions (Radu et al 2009;Bourgeois et al 2007;Schleeger et al 2009). Those reactions occur in a time scale (Fig.…”

Section: Structure and Function Of Proteinsmentioning

confidence: 99%

New opportunity to investigate physico-chemical phenomena: time-resolved X-ray and IR concurrent analysis

Marcelli

Hampai

et al. 2011

Rend. Fis. Acc. Lincei

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The characterization of both collective and individual excitations occurring in condensed matter and biological systems are of great interests for both fundamental and technological researches. New time-resolved and concurrent approaches are required to characterize complex systems where physical-chemical phenomena occur due to the interplay between localized and delocalized electrons, and in particular to characterize systems in non-equilibrium conditions. In this contribution, we present an optical layout that may take advantage of the high brilliance and the naturally wide spectral-distribution of the synchrotron radiation emission covering the range from the IR to the hard X-ray region. Thanks to the brilliance and the power associated to third generation storage rings with a conceptual new beamline design combining an IR and an X-ray beam, it will be possible to investigate dynamical processes with a time resolution down to the sub-millisecond regime, resolving plenty of correlated structural, electronic and vibrational phenomena. To demonstrate the unique advantages of this time-resolved concurrent experimental approach, among the many Invited Contribution presented to the 1st Bilateral Italian Chinese Workshop on Synchrotron radiation time resolved concurrent experiments: advantages and future applications. A new Italian route to China existing scientific cases, a few interesting but unresolved scientific cases are reviewed and potential applications of the new analytical tool are envisaged.

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“…2 This can be achieved for light activated proteins with a fast photo-cycle. On the other hand, for the investigation of noncyclic reactions the sample must be either exchanged in a flow through systems, 3 or small segments out of a larger sample area must be activated sequentially. 4 If a monochromatic light source instead of a polychromatic globar is used, the interferometer is unnecessary, and time resolution will be governed using the rise time of the detector, which is typically about 20 ns for cryogenic photovoltaic detectors.…”

Section: Introductionmentioning

confidence: 99%

A Modified Infrared Spectrometer with High Time Resolution and Its Application for Investigating Fast Conformational Changes of the GTPase Ras

2014

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Time-resolved infrared spectroscopy is a valuable tool for the investigation of proteins and protein interactions. The investigation of many biological processes is possible by means of caged compounds, which set free biologically active substances upon light activation. Some caged compounds could provide sub-nanosecond time resolution, e.g., para-hydroxyphenacyl-guanosine 5'-triphosphate (GTP) forms GTP in picoseconds. However, the time resolution in single shot experiments with rapid-scan Fourier transform infrared (FT-IR) spectrometers is limited to about 10 ms. Here we use an infrared diode laser instead of the conventional globar and achieve a time resolution of 100 ns. This allows for the time-resolved measurement of the fast Ras(off) to Ras(on) conformational change at room temperature. We quantified the activation parameters for this reaction and found that the free energy of activation for this reaction is mainly enthalpic. Investigation of the same reaction in the presence of the Ras binding domain of the effector Raf (RafRBD) reveals a four orders of magnitude faster reaction, indicating that Ras·RafRBD complex formation directly induces the conformational change. Recent developments of broadly tunable quantum cascade lasers will further improve time resolution and usability of the setup. The reported 100 ns time resolution is the best achieved for a non-repetitive experiment so far.

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Time-resolved flow-flash FT-IR difference spectroscopy: the kinetics of CO photodissociation from myoglobin revisited

Cited by 4 publications

References 0 publications

Protein dynamics observed by tunable mid-IR quantum cascade lasers across the time range from 10 ns to 1 s

Protein dynamics observed by tunable mid-IR quantum cascade lasers across the time range from 10 ns to 1 s

New opportunity to investigate physico-chemical phenomena: time-resolved X-ray and IR concurrent analysis

A Modified Infrared Spectrometer with High Time Resolution and Its Application for Investigating Fast Conformational Changes of the GTPase Ras

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