2020
DOI: 10.1038/s41586-020-2307-8
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Femtosecond-to-millisecond structural changes in a light-driven sodium pump

Abstract: Light-driven sodium pumps actively transport small cations across cellular membranes 1 .They are used by microbes to convert light into membrane potential and have become useful optogenetic tools with applications in neuroscience. While resting state structures of the prototypical sodium pump Krokinobacter eikastus rhodopsin 2 (KR2) have been solved 2,3 , it is unclear how structural alterations over time allow sodium translocation against a concentration gradient. Using the Swiss X-ray Free Electron Laser 4 ,… Show more

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Cited by 148 publications
(246 citation statements)
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“…The control map at 67 ms only contains spurious features (Figure 4A, 66 ms), which shows that a once excited jet volume entirely left the X-ray interaction region. In all previous nanosecond time-resolved crystallographic experiments on photoactive yellow protein (Jung et al, 2013;Tenboer et al, 2014), myoglobin (Schmidt et al, 2005;Srajer et al, 1996), and others (Nogly et al, 2018;Skopintsev et al, 2020), DED features are mostly localized to the chromophore and a few residues. As a consequence, the DED map sigma level is determined by the noise in the DED map as has been shown previously (Srajer et al, 2001).…”
Section: Difference Electron Density At 5 Ns and 33 Msmentioning
confidence: 92%
“…The control map at 67 ms only contains spurious features (Figure 4A, 66 ms), which shows that a once excited jet volume entirely left the X-ray interaction region. In all previous nanosecond time-resolved crystallographic experiments on photoactive yellow protein (Jung et al, 2013;Tenboer et al, 2014), myoglobin (Schmidt et al, 2005;Srajer et al, 1996), and others (Nogly et al, 2018;Skopintsev et al, 2020), DED features are mostly localized to the chromophore and a few residues. As a consequence, the DED map sigma level is determined by the noise in the DED map as has been shown previously (Srajer et al, 2001).…”
Section: Difference Electron Density At 5 Ns and 33 Msmentioning
confidence: 92%
“…Further upcoming developments relate to computing the excited state vibrational spectra of the rPSB, in relation with the very informative femtosecond data [91,111], and a possible inclusion of the dynamic polarisation of the nearest neighbour residues [15,74,75]. Unlike the here reported transient spectroscopies, ultrafast X-ray diffraction has the potential of revealing the detailed dynamic structural and electronic changes of the rPSB and its nearest environment, provided these experiments be carried out under single photon excitation conditions [63,72,73,126]. Beyond the excited state lifetimes and vibrational coherences, another central property is the reaction quantum yield (IQY), for which we still have to understand and learn the lessons that the outstanding high values found in Rho and bR can teach us, when these are compared with the modest values obtained for the biomimetic switches in solution [20] or the low values (<2%) observed for the crowed alkenes designed as molecular rotary machine [26].…”
Section: Towards a Rational Prediction Of The Excited State Lifetimentioning
confidence: 97%
“…Its crystallographic structure was resolved [61,62] [61]. The overall structural changes associated with Na + pumping were very recently analyzed by time-resolved serial X-ray crystallography and QM/MM simulations, covering the pico-to milli-second time scales [63]. However, the changes in the SB-D116 distance were observed to be much smaller than expected, and no clear evidence for the existence of two rotameric states was found in the photocycle at pH=8.0.…”
Section: B Effects Of Mutations and Ph On Excited State Dynamics Of mentioning
confidence: 99%
“…While at first, it was modeled as an unbranched unidirectional photocycle with five distinct photointermediates, 14 later on more complex models were proposed. 15,16,17 KR2's photocycle has already been studied extensively with various techniques [18][19][20][21][22][23] but still remains to be understood completely. Especially the spectral location of the photointermediates, as well as the equilibria within the photocycle remain elusive.…”
Section: Introductionmentioning
confidence: 99%