期刊
ANALYTICAL CHEMISTRY
卷 90, 期 17, 页码 10494-10500出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.8b02531
关键词
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资金
- Heisenberg fellowship [K03580/4-2]
- DFG [K03580/5-1]
- Swiss National Science Foundation [200020_178942]
- Swiss National Science Foundation (SNF) [200020_178942] Funding Source: Swiss National Science Foundation (SNF)
The kinetic analysis of irreversible protein reactions requires an analytical technique that provides access to time-dependent infrared spectra in a single shot. Here, we present a spectrometer based on dual-frequency-comb spectroscopy using mid-infrared frequency combs generated by quantum cascade lasers. Attenuation of the intensity of the combs by molecular vibrational resonances results in absorption spectra covering 55 cm(-1) in the fingerprint region. The setup has a native resolution of 0.3 cm(-1), noise levels in the mu OD range, and achieves sub-microsecond time resolution. We demonstrate the simultaneous recording of both spectra and transients of the photoactivated proton pump bacteriorhodopsin. More importantly, a single shot, i.e., a single visible light excitation, is sufficient to extract spectral and kinetic characteristics of several intermediates in the bacteriorhodopsin photocycle. This development paves the way for the noninvasive analysis of enzymatic conversions with high time resolution, broad spectral coverage, and minimal sample consumption.
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