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1 DOI: /NCHEM.2398 Locl virtionl coherences drive the primry photochemistry of vision Philip J. M. Johnson 1, Alexei Hlpin 1, Tkefumi Morizumi 2, Vlentyn I. Prokhorenko 3, Oliver P. Ernst 2,4, & R. J. Dwyne Miller 1,3 1 Institute for Opticl Sciences nd Deprtments of Chemistry nd Physics, University of Toronto, 80 St. George Street, Toronto, Ontrio M5S 3H6, Cnd. 2 Deprtment of Biochemistry, University of Toronto, 1 King s College Circle, Toronto, Ontrio M5S 1A8, Cnd. 3 Mx Plnck Institute for the Structure nd Dynmics of Mtter, Atomiclly Resolved Dynmics Division, Building 99 (CFEL), Luruper Chussee 149, Hmurg, Germny. 4 Deprtment of Moleculr Genetics, University of Toronto, 1 King s College Circle, Toronto, Ontrio M5S 1A8, Cnd. NATURE CHEMISTRY 1
2 1 Pulse chrcteriztion To temporlly compress the output of the NOPA, we employed comintion of deformle mirror nd prism compressors. The deformle mirror compressor [1, 2] consisted of 600 grooves/mm grting lzed for 500 nm, n f = 300 mm sphericl mirror, nd memrne-sed 13 3 rry deformle mirror (OKO Technologies) ll in 4f configurtion, while the folded prism compressor consisted of two equilterl fused silic prisms seprted y 90 cm. Trnsient grting FROG [3] ws collected in situ t the smple position nd nlysed with the commercil FROG3 softwre (Femtosecond Technologies, USA). The optimlly compressed pulse with the clenest temporl profile (i.e., without stellites or significnt mplitude in the wings of the min pulse) is shown in Fig. S1, where the temporl fwhm of the intensity profile is 11 fs. 2 Decy-ssocited spectr Glol nlysis [4] of the spectrlly-resolved trnsient grting dt ws performed s descried previously [5]. Three distinct components were recovered y this nlysis (see π -π - Figure S1: Trnsient grting FROG nlysis of the ultrshort pulses used for trnsient grting mesurements of rhodopsin., A countour plot of the trnsient grting FROG signl of the optimlly compressed pulses used in the experiment. Contours re drwn t 10% intervls eginning t 5% of the signl mximum., The recovered intensity profile (solid line) nd the ssocited phse (dshed line) of the ultrshort pulse in. NATURE CHEMISTRY 2
3 c fs 106 fs 2.1 ps Figure S2: Decy-ssocited spectrl nlysis of the trnsient grting response of rhodopsin., The trnsient grting response of rhodopsin over [0.01, 1.5] ps., Glol fitting of the trnsient grting dt required three kinetic components to ccurtely reconstruct the oserved dynmics. c, The three decy-ssocited spectr recovered from the glol nlysis revel su-50 fs, 100 fs, nd 2.1 ps kinetic components, ttriutle to leching of the ground stte nd photoproduct formtion, ground stte recovery, nd the thermlized thorhodopsin formtion, respectively. Fig. S2). First, su-50 fs component (Fig. S2c, ornge trce) comprised of lech response t the ground stte liner sorption frequencies (spnning cm 1 ) nd photoproduct sorption t the differentil spectrl coordintes ssocited with the thorhodopsin intermedite (in the vicinity of cm 1 ). Second, 100 fs component (Fig. S2c, green trce) comprised lmost strictly of recovery of the ground stte lech, in concurrence with the oservtions for the single frequency trces descried in the min text. Finlly, 2 ps component is lso recovered (Fig. S2c, lue trce), ttriutle to the thermlized thorhodopsin intermedite, nlogous to the similr component oserved in cteriorhodopsin ttriuted to the K intermedite [5, 6] which displys similr kinetic timescle nd spectrl profile. 3 Virtionl filtering nd fitting To determine the virtionl dynmics of the retinl chromophore during isomeriztion, the popultion-sutrcted dynmics were filtered y tpered cosine window in the NATURE CHEMISTRY 3
4 Figure S3: The experimentl time-domin virtionl trnsients (solid coloured lines), the reconstructed virtionl trnsients following the ppliction of five Fourier filters (superimposed dshed lck lines) nd the difference etween the two (solid lck lines) for, ν proe = cm 1 nd, ν proe = cm 1. Fourier domin to isolte virtionl frequency rnges of interest, nd susequently inverse Fourier trnsformed to recover the time-domin response of suset of ll of the virtionl modes. To confirm the completeness of the filtering, comprison etween the unfiltered time-domin virtionl trnsients nd the recovered virtionl response following the ppliction of ll five Fourier filters is shown in Fig. S3, where the solid lck lines show the difference etween the popultion-sutrcted dt nd the sum of the filtered response, showing excellent greement for ll ut the very initil time points for oth proe regions of interest. Figure S4: Virtionl filtering nd fitting., The complete virtionl response t ν proe = cm 1 (solid red line) nd the result of pplying Fourier filter to isolte the low frequency deloclized torsionl modes (solid lck line). Inset: the power spectrum of the virtionl modes (solid red line) nd the Fourier-domin filter (solid lck line)., Time domin fitting (dshed red line) of the filtered virtionl trnsients (solid lck line, s in ) nd the residuls of the fit (dshed lck line, elow), for eight decying sinusoids. NATURE CHEMISTRY 4
5 Ech individully filtered trce (s in Fig. 3 of the min text) represents suset of the totl virtionl response, nd is thus fr more menle to time-domin fitting. An exmple of this process is shown in Fig. S4 for the virtionl response t ν proe = cm 1 with the lowest frequency Fourier filter pplied to resolve the deloclized torsionl response of the retinl chromophore. The inverse Fourier trnsformed dt shows only the low frequency response, nd ws successfully fit (Fig. S4) y sum of decying sinusoids, resulting in the recovery of eight different virtionl modes with frequencies etween cm 1 (see Tle 1 nd Fig. S5). 4 Stick spectrum The result of the complete nlysis of virtionl trnsients is displyed in Tle 1 of the min text, nd here we disply (see Fig. S5) stick spectrum of the mplitudes of the vrious modes resolved y time-domin nlysis of the filtered virtionl trnsients. Nrrow sticks represent ground stte modes with dephsing times 200 fs, while excited stte modes with dephsing times 100 fs re shown s thicker lines. Sticks in lue represent virtions recovered t the lech nd, red sticks represent virtions recovered t the photoproduct nd. Figure S5: Stick spectrum of the modes outlined in Tle 1 of the min text. Thin lines represent long-dephsing time virtions which cn e ttriuted to ground stte virtions. Shorter dephsing time modes ( 100 fs) re shown s thick lines, nd re ttriuted excited stte nucler motions. NATURE CHEMISTRY 5
6 References [1] MR Armstrong, P Plcht, EA Ponomrev, nd RJD Miller. Verstile 7-fs opticl prmetric pulse genertion nd compression y use of dptive optics. Opt. Lett. 26, (2001). [2] A Bltušk, T Fuji, nd T Koyshi. Visile pulse compression to 4 fs y opticl prmetric mplifiction nd progrmmle dispersion control. Opt. Lett. 27, (2002). [3] R Treino, KW DeLong, DN Fittinghoff, JN Sweetser, MA Krumügel, BA Richmn. Mesuring ultrshort lser pulses in the time-frequency domin using frequency-resolved opticl gting. Rev. Sci. Instruments 68, (1997). [4] IHM vn Stokkum, DS Lrsen, nd R vn Grondelle. Glol nd trget nlysis of time-resolved spectr. Biochimic et Biophysic Act 1657, (2004). [5] VI Prokhorenko, AM Ngy, LS Brown, nd RJD Miller. On the mechnism of wekfield coherent control of retinl isomeriztion in cteriorhodopsin. Chem. Phys. 341, (2007). [6] PJM Johnson, A Hlpin, T Morizumi, LS Brown, VI Prokhorenko, OP Ernst, nd RJD Miller. The photocycle nd ultrfst virtionl dynmics of cteriorhodopsin in lipid nnodiscs. Phys. Chem. Chem. Phys. 16, (2014). NATURE CHEMISTRY 6
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