Spectroscopic Studies of Model Photo-receptors: Validation of a Nanosecond Time-resolved Micro-spectrophotometer Design Using Photoactive Yellow Protein and α-phycoerythrocyanin

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2013 Sep 26

PMID 24065094

Citations 6

Authors

Namrta Purwar

Jason Tenboer

Shailesh Tripathi

Marius Schmidt

Affiliations

Soon will be listed here.

Abstract

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 α-phycoerythrocyanin (α-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.

Citing Articles

Confinement in crystal lattice alters entire photocycle pathway of the Photoactive Yellow Protein.

Konold P, Arik E, Weissenborn J, Arents J, Hellingwerf K, van Stokkum I Nat Commun. 2020; 11(1):4248.

PMID: 32843623 PMC: 7447820. DOI: 10.1038/s41467-020-18065-9.

Watching Proteins Function with Time-resolved X-ray Crystallography.

Srajer V, Schmidt M J Phys D Appl Phys. 2018; 50(37).

PMID: 29353938 PMC: 5771432. DOI: 10.1088/1361-6463/aa7d32.

A short history of structure based research on the photocycle of photoactive yellow protein.

Schmidt M Struct Dyn. 2017; 4(3):032201.

PMID: 28191482 PMC: 5291790. DOI: 10.1063/1.4974172.

Femtosecond structural dynamics drives the trans/cis isomerization in photoactive yellow protein.

Pande K, Hutchison C, Groenhof G, Aquila A, Robinson J, Tenboer J Science. 2016; 352(6286):725-9.

PMID: 27151871 PMC: 5291079. DOI: 10.1126/science.aad5081.

Time-resolved serial crystallography captures high-resolution intermediates of photoactive yellow protein.

Tenboer J, Basu S, Zatsepin N, Pande K, Milathianaki D, Frank M Science. 2014; 346(6214):1242-6.

PMID: 25477465 PMC: 4361027. DOI: 10.1126/science.1259357.

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