Electric-field dependent decays of two spectroscopically different M-states of photosensory rhodopsin II from Natronobacterium pharaonis

Biophysical Journal
Laura RivasPeter Hildebrandt

Abstract

Sensory rhodopsin II (NpSRII) from Natronobacterium pharaonis was studied by resonance Raman (RR) spectroscopic techniques. Using gated 413-nm excitation, time-resolved RR measurements of the solubilized photoreceptor were carried out to probe the photocycle intermediates that are formed in the submillisecond time range. For the first time, two M-like intermediates were identified on the basis of their C=C stretching bands at 1568 and 1583 cm(-1), corresponding to the early M(L)(400) state with a lifetime of 30 micro s and the subsequent M(1)(400) state with a lifetime of 2 ms, respectively. The unusually high C=C stretching frequency of M(1)(400) has been attributed to an unprotonated retinal Schiff base in a largely hydrophobic environment, implying that the M(L)(400) --> M(1)(400) transition is associated with protein structural changes in the vicinity of the chromophore binding pocket. Time-resolved surface enhanced resonance Raman experiments of NpSRII electrostatically bound onto a rotating Ag electrode reveal that the photoreceptor runs through the photocycle also in the immobilized state. Surface enhanced resonance Raman spectra are very similar to the RR spectra of the solubilized protein, ruling out adsorption-induced...Continue Reading

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Citations

Aug 23, 2008·Proceedings of the National Academy of Sciences of the United States of America·X JiangJ Heberle
Nov 12, 2013·Biochimica Et Biophysica Acta·Víctor A Lórenz-Fonfría, Joachim Heberle
Apr 28, 2004·Biophysical Journal·Yukako IkeuraNaoki Kamo
Dec 1, 2006·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Hendrik NaumannDaniel H Murgida
Dec 23, 2004·Accounts of Chemical Research·Daniel H Murgida, Peter Hildebrandt
Aug 5, 2010·Journal of the American Chemical Society·Xiue JiangJoachim Heberle
Sep 30, 2005·Journal of the American Chemical Society·Smilja TodorovicDaniel H Murgida

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