Carino Ferrante Advisor: Tullio ScopignoPontecorvo , C. Ferrante Elles and TS, Optics Express 21,...
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Ultrafast photoinduced dynamics in heme proteins Carino Ferrante Advisor: Tullio Scopigno Dipartimento di Fisica - University of Rome “Sapienza” PhD XXVI
Transcript of Carino Ferrante Advisor: Tullio ScopignoPontecorvo , C. Ferrante Elles and TS, Optics Express 21,...
Ultrafast photoinduced dynamics in heme proteins
Carino Ferrante
Advisor: Tullio Scopigno
Dipartimento di Fisica - University of Rome “Sapienza”
PhD XXVI
The Pump-Probe approach…
Ultrafast Dynamics with structural sensitivity:
• Time Resolved Resonance Raman (TR3)
Photoproduct
Reactant
Ultrafast Dynamics with structural sensitivity:
• Time Resolved Resonance Raman (TR3)
CW
δω=30cm-1 δt=0.7ps
Jean-Louis Martin et al. 2007
“…However, such studies are extremely challenging for the
physiologically relevant ligand oxygen and have not been
reported yet, even for the well studied oxygen carriers myoglobin
and hemoglobin, presumably at least in part because of technical
difficulties”
“…Because of the decreased spectral resolution in the
subpicosecond Raman experiments, lines in these spectra are
broader than in the steady-state spectra obtained with cw
excitation.”
Heisenberg uncertainty principle δωδt ≥ 15 ps cm-1
Ultrafast Dynamics with structural sensitivity:
• Time Resolved Resonance Raman (TR3)
CW
δω=30cm-1 δt=0.7ps
Jean-Louis Martin et al. 2007
“…However, such studies are extremely challenging for the
physiologically relevant ligand oxygen and have not been
reported yet, even for the well studied oxygen carriers myoglobin
and hemoglobin, presumably at least in part because of technical
difficulties”
“…Because of the decreased spectral resolution in the
subpicosecond Raman experiments, lines in these spectra are
broader than in the steady-state spectra obtained with cw
excitation.”
Heisenberg uncertainty principle δωδt ≥ 15 ps cm-1 Instrument limited
kinetics
“transform limited” spontaneous Raman
δωδt>15ps·cm-1
Ram
an
Pu
lse
800 850 900 950
(nm)
Raman
Pulse
Raman
Pulse
Timeline
FSRS in a nutshell:
BB stimulated Raman
Ram
an
Pu
lse
Ram
an
Pro
be
800 850 900 950
(nm)
Raman
Pulse
Pump on
Raman
Probe
Raman
Pulse
Timeline
Raman
Probe
FSRS in a nutshell:
Ph
oto
chem
ical
Pu
mp
Ram
an
Pu
lse
Ram
an
Pro
be
En
erg
y
“Reaction” coordinate
800 850 900 950
(nm)
Raman
Pulse
Pump on
Raman
Probe
Photochemical
Pump
∆t
Raman
Pulse
Timeline
Raman
Probe
Unrestricted
time precision
FSRS in a nutshell:
FSRS Setup Timing cycle – 1 ms timebase:
WLC ON ON ON ON ….
RAMAN PULSE ON OFF ON OFF ….
PHOTOCHEM PUMP ON ON OFF OFF ….
E. Pontecorvo, M.S. Kapetanaki, M. Badioli, D. Brida, M. Marangoni G. Cerullo and TS, Optics Express 19, 1107 (2011)
FSRS Setup Timing cycle – 1 ms timebase:
WLC ON ON ON ON ….
RAMAN PULSE ON OFF ON OFF ….
PHOTOCHEM PUMP ON ON OFF OFF ….
E. Pontecorvo, M.S. Kapetanaki, M. Badioli, D. Brida, M. Marangoni G. Cerullo and TS, Optics Express 19, 1107 (2011)
WLG
WL
Prob
e
FSRS Setup Timing cycle – 1 ms timebase:
WLC ON ON ON ON ….
RAMAN PULSE ON OFF ON OFF ….
PHOTOCHEM PUMP ON ON OFF OFF ….
E. Pontecorvo, M.S. Kapetanaki, M. Badioli, D. Brida, M. Marangoni G. Cerullo and TS, Optics Express 19, 1107 (2011)
Raman Pulse
Ram
an
Pu
mp
FSRS Setup Timing cycle – 1 ms timebase:
WLC ON ON ON ON ….
RAMAN PULSE ON OFF ON OFF ….
PHOTOCHEM PUMP ON ON OFF OFF ….
E. Pontecorvo, M.S. Kapetanaki, M. Badioli, D. Brida, M. Marangoni G. Cerullo and TS, Optics Express 19, 1107 (2011)
Raman Pulse
Ram
an
Pu
mp
E. Pontecorvo, C. Ferrante, C. Elles and TS, Optics Express 21, 6866 (2013)
FSRS Setup Timing cycle – 1 ms timebase:
WLC ON ON ON ON ….
RAMAN PULSE ON OFF ON OFF ….
PHOTOCHEM PUMP ON ON OFF OFF ….
E. Pontecorvo, M.S. Kapetanaki, M. Badioli, D. Brida, M. Marangoni G. Cerullo and TS, Optics Express 19, 1107 (2011)
Raman Pulse
Ram
an
Pu
mp
E. Pontecorvo, C. Ferrante, C. Elles and TS, Optics Express 21, 6866 (2013)
01234
t = 0.25 ps
t = 0 ps
10
3 R
aman
Gai
n
Energy (cm-1)
01234
01234
01234
01234
01234
01234
01234
250 500 750 1000 1250 150001234
t = 30 ps
t = 5.0 ps
t = 3.0 ps
t = 1.5 ps
t = 0.40 ps
t = 0.30 ps
t = -10 ps
Mb deOxy
FSRRS
FSRS at work: sub-ps in Mb
TR3
CW Δ𝒕 = 𝟐. 𝟓 𝒑𝒔
01234
t = 0.25 ps
t = 0 ps
10
3 R
aman
Gai
n
Energy (cm-1)
01234
01234
01234
01234
01234
01234
01234
250 500 750 1000 1250 150001234
t = 30 ps
t = 5.0 ps
t = 3.0 ps
t = 1.5 ps
t = 0.40 ps
t = 0.30 ps
t = -10 ps
Mb deOxy
FSRS at work: sub-ps in Mb
τ~5.2ps
Equilibrium DeOxy
Analysis of the ν4
Mb deOxy
MbCO
τ~5.2ps
HbCO
HOT Hb
Photoexcited Myoglobin
What happens after photolysis (t<50fs)?
1. Existence of electronic intermediate states
HbCO
HOT Hb
Photoexcited Myoglobin
What happens after photolysis (t<50fs)?
1. Existence of electronic intermediate states
2. Vibrational cooling from a hot ground state
Hypothesis n°1 Existence of electronic states
430 440 450 460 4700
1
Soret absorption in DeOxy Mb
Virtual level
Electronic level
Δ𝜔𝑀𝑏 MbI MbII
Mb+
Mb
Hypothesis n°1 Existence of electronic states
430 440 450 460 4700
1
Soret absorption in DeOxy Mb
Virtual level
Electronic level
Δ𝜔𝑀𝑏 MbI MbII
Mb+
Mb
Hypothesis n°1 Existence of electronic states
430 440 450 460 4700
1
Soret absorption in DeOxy Mb
Virtual level
Electronic level
Δ𝜔𝑀𝑏𝐼
MbI MbII
Mb+
Mb
Hypothesis n°1 Existence of electronic states
No new peaks
Mbdeoxy RP @ 460 nm
HbCO
Unlikely existence of
electronic intermediate states
Conclusions
• Simple setup for FSRRS allows accessing molecular resonances in a wide
wavelength range.
• Mb: FSRRS suggests vibrational relaxation as opposed to intermediate states, elucidating the energy redistribution pathway
sub-ps thermometer
• Ngb: Similar vibrational relaxation with different anharmonicity factor and time scale
PhD activities
1. E. Pontecorvo, C. Ferrante, M. Ferretti, M. Ortolani, D. Polli, G. Ruocco, G. Cerullo and T. Scopigno. Applied Physics Letters, 98:246102, 2011.
2. E. Pontecorvo, C. Ferrante, C. G. Elles, and T. Scopigno. Opt. Express, 21(6):6866, 2013.
3. C. Ferrante, E. Pontecorvo, G. Cerullo, A. Chiasera, G. Ruocco, W. Schirmacher, and T. Scopigno. Nature Communications, 04:1793, 2013.
...Further work in progress
