期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 14, 期 9, 页码 18881-18898出版社
MDPI
DOI: 10.3390/ijms140918881
关键词
nanosecond spectroscopy; time-resolved spectroscopy; photoreceptor proteins
资金
- [NSF-0952643]
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [0952643] Funding Source: National Science Foundation
Time-resolved spectroscopic experiments have been performed with protein in solution and in crystalline form using a newly designed microspectrophotometer. The time-resolution of these experiments can be as good as two nanoseconds (ns), which is the minimal response time of the image intensifier used. With the current setup, the effective time-resolution is about seven ns, determined mainly by the pulse duration of the nanosecond laser. The amount of protein required is small, on the order of 100 nanograms. Bleaching, which is an undesirable effect common to photoreceptor proteins, is minimized by using a millisecond shutter to avoid extensive exposure to the probing light. We investigate two model photoreceptors, photoactive yellow protein (PYP), and -phycoerythrocyanin (-PEC), on different time scales and at different temperatures. Relaxation times obtained from kinetic time-series of difference absorption spectra collected from PYP are consistent with previous results. The comparison with these results validates the capability of this spectrophotometer to deliver high quality time-resolved absorption spectra.
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