Selective Tautomer Production and Cryogenic Ion Spectroscopy of Radical Cations: The Uracil and Thymine Cases.

The vibrational and electronic spectroscopy of the radical cations of two nucleobases (NB) (uracil and thymine) was studied by cryogenic ion photodissociation spectroscopy. The radical cations have been generated from the photodissociation of NB-Ag+ complexes. A charge transfer process from the NB to Ag+ governs the deactivation mechanism, leading to the formation of the radical cation without further tautomerization. Single- and double-resonance spectroscopy allows for structural assignments of both the silver complexes and the radical cations by comparison with DFT-based calculations. Interestingly, a tautomer-dependent fragmentation is observed in the thymine enol form that involves the loss of NCO, a fragment which was never reported before for this NB. This selective photodissociation of silver complexes containing aromatic chromophore greatly expands the current technique to produce isomer-selected radical cations in the gas phase providing benchmark experimental data to assess calculations of open-shell species.

Conformer-selective Photodynamics of TrpH+ -H2 O.

The photodynamics of protonated tryptophan and its mono hydrated complex TrpH+ -H2 O has been revisited. A combination of steady-state IR and UV cryogenic ion spectroscopies with picosecond pump-probe photodissociation experiments sheds new lights on the deactivation processes of TrpH+ and conformer-selected TrpH+ -H2 O complex, supported by quantum chemistry calculations at the DFT and coupled-cluster levels for the ground and excited states, respectively. TrpH+ excited at the band origin exhibits a transient of less than 100 ps, assigned to the lifetime of the excited state proton transfer (ESPT) structure. The two experimentally observed conformers of TrpH+ -H2 O have been assigned. A striking result arises from the conformer-selective photodynamics of TrpH+ -H2 O, in which a single water molecule inserted in between the ammonium and the indole ring hinders the barrierless ESPT reaction responsible for the ultra-fast deactivation process observed in the other conformer and in bare TrpH+ .

Cryogenic IR and UV spectroscopy of isomer-selected cytosine radical cation.

Oxidation of the nucleobases is of great concern for the stability of DNA strands and is considered as a source of mutagenesis and cancer. However, precise spectroscopy data, in particular in their electronic excited states are scarce if not missing. We here report an original way to produce isomer-selected radical cations of DNA bases, exemplified in the case of cytosine, through the photodissociation of cold cytosine-silver (C-Ag+) complex. IR-UV dip spectroscopy of C-Ag+ features fingerprint bands for the two keto-amino cytosine tautomers. UV photodissociation (UVPD) of the isomer-selected C-Ag+ complexes produces the cytosine radical cation (C˙+) without isomerization. IR-UV cryogenic ion spectroscopy of C˙+ allows for the unambiguous structural assignment of the two keto-amino isomers of C˙+. UVPD spectroscopy of the isomer-selected C˙+ species is recorded at a unique spectral resolution. These benchmark spectroscopic data of the electronic excited states of C˙+ are used to assess the quantum chemistry calculations performed at the TD-DFT, CASSCF/CASPT2 and CASSCF/MRCI-F12 levels.

Excited state dynamics of protonated dopamine: hydration and conformation effects.

Electronic and vibrational spectroscopy in a cryogenic ion trap has been applied to protonated dopamine water clusters and assigned with the help of quantum chemistry calculations performed in the ground and electronic excited states. A dramatic hydration effect is observed when dopamine is solvated by three water molecules. The broad electronic spectra recorded for the bare and small water clusters containing protonated dopamine turn to sharp, well-resolved vibronic transitions in the 1-3 complex. This reflects the change induced by hydration in the photodynamics of protonated dopamine which is initially controlled by an excited state proton transfer (ESPT) reaction from the ammonium group toward the catechol ring. Interestingly, conformer selectivity is revealed in the 1-3 complex which shows two low lying energy conformers for which the ESPT reaction is prevented or not depending on the H-bond network formed between the dopamine and water molecules.

Hydration-controlled excited-state relaxation in protonated dopamine studied by cryogenic ion spectroscopy.

Ultraviolet (UV) and infrared (IR) spectra of protonated dopamine (DAH+) and its hydrated clusters DAH+(H2O)1-3 are measured by cryogenic ion spectroscopy. DAH+ monomer and hydrated clusters with up to two water molecules show a broad UV spectrum, while it turns to a sharp, well-resolved one for DAH+-(H2O)3. Excited state calculations of DAH+(H2O)3 reproduce these spectral features. The conformer-selected IR spectrum of DAH+(H2O)3 is measured by IR dip spectroscopy, and its structure is assigned with the help of quantum chemical calculations. The excited state lifetime of DAH+ is much shorter than 20 ps, the cross correlation of the ps lasers, revealing a fast relaxation dynamics. The minimal energy path along the NH → π proton transfer coordinate exhibits a low energy barrier in the monomer, while this path is blocked by the high energy barrier in DAH+(H2O)3. It is concluded that the excited state proton transfer in DAH+ is inhibited by water-insertion.

Excited-state proton transfer in protonated adrenaline revealed by cryogenic UV photodissociation spectroscopy.

We report a comprehensive study of the structures and deactivation processes of protonated adrenaline through cryogenic UV photodissociation spectroscopy. Single UV and double-resonance UV-UV hole burning spectroscopies have been performed and compared to coupled-cluster SCS-CC2 calculations performed on the ground and first electronic states. Three conformers were assigned, two lowest energy gauche conformers along with a higher energy conformer with an extended structure which is indeed the global minimum in solution. This demonstrates the kinetic trapping of this high energy gas phase conformer during the electrospray process. At the band origin of all conformers, the main fragmentation channel is the Cα-Cß bond cleavage, triggered by an excited state proton transfer to the catechol ring. Internal conversion leading to the water loss channel competes with the direct dissociation and tends to prevail with the increase of excess energy brought by the UV laser. Picosecond time-resolved pump-probe spectroscopy was performed to measure the excited state lifetimes of the three conformers of AdH+, which decay with the increase of excess energy in the ππ* state, from 2 ns at the band origin down to few hundreds of picoseconds 0.5 eV to the blue. Finally, about 0.8 eV above the band origin, the πσ* state is directly reached, leading to the opening of the H-loss channel.

UV Photoinduced Dynamics of Conformer-Resolved Aromatic Peptides.

A detailed understanding of radiative and nonradiative processes in peptides containing an aromatic chromophore requires the knowledge of the nature and energy level of low-lying excited states that could be coupled to the bright 1ππ* excited state. Isolated aromatic amino acids and short peptides provide benchmark cases to study, at the molecular level, the photoinduced processes that govern their excited state dynamics. Recent advances in gas phase laser spectroscopy of conformer-selected peptides have paved the way to a better, yet not fully complete, understanding of the influence of intramolecular interactions on the properties of aromatic chromophores. This review aims at providing an overview of the photophysics and photochemistry at play in neutral and charged aromatic chromophore containing peptides, with a particular emphasis on the charge (electron, proton) and energy transfer processes. A significant impact is exerted by the experimental progress in energy- and time-resolved spectroscopy of protonated species, which leads to a growing demand for theoretical supports to accurately describe their excited state properties.

Water binding to FeIII hemes studied in a cooled ion trap: characterization of a strong 'weak' ligand.

The interaction of a water molecule with ferric heme-iron protoporphyrin ([PP FeIII]+) has been investigated in the gas phase in an ion trap and studied theoretically by density functional theory. It is found that the interaction of water with ferric heme leads to a stable [PP-FeIII-H2O]+ complex in the intermediate spin state (S = 3/2), in the same state as its unligated [PP-FeIII]+ homologue, without spin crossing during water attachment. Using the Van't Hoff equation, the reaction enthalpy for the formation of a Fe-OH2 bond has been determined for [PP-FeIII-H2O]+ and [PP-FeIII-(H2O)2]+. The corrected binding energy for a single Fe-H2O bond is -12.2 ± 0.6 kcal mol-1, while DFT calculations at the OPBE level yield -11.7 kcal mol-1. The binding energy of the second ligation yielding a six coordinated FeIII atom is decreased with a bond energy of -9 ± 0.9 kcal mol-1, well reproduced by calculations as -7.1 kcal mol-1. However, calculations reveal features of a weaker bond type, such as a rather long Fe-O bond with 2.28 Å for the [PP-FeIII-H2O]+ complex and the absence of a spin change by complexation. Thus despite a strong bond with H2O, the FeIII atom does not show, through theoretical modelling, a strong acceptor character in its half filled 3dz2 orbital. It is also observed that the binding properties of H2O to hemes seem strikingly specific to ferric heme and we have shown, experimentally and theoretically, that the affinity of H2O for protonated heme [H PP-Fe]+, an intermediate between FeIII and FeII, is strongly reduced compared to that for ferric heme.

UV Photofragmentation of Cold Cytosine-M+ Complexes (M+: Na+, K+, Ag+).

The UV photofragmentation spectra of cold cytosine-M+ complexes (M+: Na+, K+, Ag+) were recorded and analyzed through comparison with geometry optimizations and frequency calculations of the ground and excited states at the SCS-CC2/Def2-SVPD level of theory. While in all complexes, the ground state minimum geometry is planar (Cs symmetry), the ππ* state minimum geometry has the NH2 group slightly twisted and an out-of-plane metal cation. This was confirmed by comparing the simulated ππ* Franck-Condon spectra with the vibrationally resolved photofragmentation spectra of CytNa+ and CytK+. Vertical excitation transitions were also calculated to evaluate the energies of the CT states involving the transfer of an electron from the Cyt moiety to M+. For both CytK+ and CytNa+ complexes, the first CT state corresponds to an electron transfer from the cytosine aromatic π ring to the antibonding σ* orbital centered on the alkali cation. This πσ* state is predicted to lie much higher in energy (>6 eV) than the band origin of the π-π* electronic transition (around 4.3 eV) unlike what is observed for the CytAg+ complex for which the first excited state has a nOσ* electronic configuration. This is the reason for the absence of the Cyt+ + M charge transfer fragmentation channel for CytK+ and CytNa+ complexes.

The dramatic effect of N-methylimidazole on trans axial ligand binding to ferric heme: experiment and theory.

The binding energy of CO, O2 and NO to isolated ferric heme, [FeIIIP]+, was studied in the presence and absence of a σ donor (N-methylimidazole and histidine) as the trans axial ligand. This study combines the experimental determination of binding enthalpies by equilibrium measurements in a low temperature ion trap using the van't Hoff equation and high level DFT calculations. It was found that the presence of N-methylimidazole as the axial ligand on the [FeIIIP]+ porphyrin dramatically weakens the [FeIIIP-ligand]+ bond with an up to sevenfold decrease in binding energy owing to the σ donation by N-methylimidazole to the FeIII(3d) orbitals. This trans σ donor effect is characteristic of ligation to iron in hemes in both ferrous and ferric redox forms; however, to date, this has not been observed for ferric heme.

Molecular Recognition by a Short Partial Peptide of the Adrenergic Receptor: A Bottom-Up Approach.

Receptor-neurotransmitter molecular recognition is key for neurotransmission. Although crystal structures of the receptors are known, the mechanism for recognition is not clear. Reported here is the ultraviolet (UV) and infrared (IR) spectra of complexes between a partial peptide (SIVSF), mimicking the binding motif of a catechol ring in the adrenergic receptor, and various ligands. The UV spectra show that two isomers coexist in the complex of SIVSF with properly recognized ligands, such as protonated adrenaline (adrenalineH+ ). From IR spectra, they are assigned to catechol- and amino-bound structures. The catechol-bound structure is not observed when the ligand is replaced by nonproper molecules, such as noradrenalineH+ . The results suggest that SIVSF not only recognizes the catechol ring but can distinguish differences in the amine side chain. The method provides a new possibility for screening molecules as potential therapeutics for activating the receptor.

Tautomerism and electronic spectroscopy of protonated 1- and 2-aminonaphthalene.

Experimental and theoretical investigations of the excited states of protonated 1- and 2-aminonaphthalene are presented. The electronic spectra are obtained by laser induced photofragmentation of the ions captured in a cold ion trap. Using ab initio calculations, the electronic spectra can be assigned to different tautomers which have the proton on the amino group or on the naphthalene moiety. It is shown that the tautomer distribution can be varied by changing the electrospray source conditions, favoring either the most stable form in solution (amino protonation) or that in the gas phase (aromatic ring protonation). Calculations for larger amino-polyaromatics predict that these systems should behave as "proton sponges" i.e. have a proton affinity larger than 11 eV.

Excited State Dynamics of Cold Protonated Cytosine Tautomers: Characterization of Charge Transfer, Intersystem Crossing, and Internal Conversion Processes.

Charge transfer reactions are ubiquitous in chemical reactivity and often viewed as ultrafast processes. For DNA, femtochemistry has undeniably revealed the primary stage of the deactivation dynamics of the locally excited state following electronic excitation. We here demonstrate that the full time scale excited state dynamics can be followed up to milliseconds through an original pump-probe photodissociation scheme applied to cryogenic ion spectroscopy. Protonated cytosine is chosen as a benchmark system in which the locally excited 1ππ* state decays in the femtosecond range toward long-lived charge transfer and triplet states with lifetimes ranging from microseconds to milliseconds, respectively. A three-step mechanism (1ππ* → 1CT → 3ππ*) is proposed where internal conversion from each state can occur leading ultimately to fragmentation in the ground electronic state.

Dioxygen Binding to Protonated Heme in the Gas Phase, an Intermediate Between Ferric and Ferrous Heme.

With a view to characterizing the influence of the electronic structure of the Fe atom on the nature of its bond with dioxygen (O2 ) in heme compounds, a study of the UV/Vis action spectra and binding energies of heme-O2 molecules has been undertaken in the gas phase. The binding reaction of protonated ferrous heme [FeII -hemeH]+ with O2 has been studied in the gas phase by determining the equilibrium of complexed [FeII -hemeH(O2 )]+ with uncomplexed protonated heme in an ion trap at controlled temperatures. The binding energy of O2 to the Fe atom of protonated ferrous heme was obtained from a van't Hoff plot. Surprisingly, this energy (1540±170 cm-1 , 18.4±2 kJ mol-1 ) is intermediate between those of ferric heme and ferrous heme. This result is interpreted in terms of a delocalization of the positive charge over the porphyrin cycle, such that the Fe atom bears a fractional positive charge. The resulting electron distribution on the Fe atom differs notably from that of a purely low-spin ferrous heme [FeII -heme(O2 )] complex, as deduced from its absorption spectrum. It also differs from that of ferric heme [FeIII -heme(O2 )]+ , as evidenced by the absorption spectra. Protonated heme creates a specific bond that cannot accommodate strong σ donation.

Multiscale excited state lifetimes of protonated dimethyl aminopyridines.

The excited state dynamics of protonated ortho (2-) and para (4-) dimethyl aminopyridine molecules (DMAPH(+)) has been studied through pump-probe photofragmentation spectroscopy and excited state coupled-cluster CC2 calculations. Multiscale temporal dynamics has been recorded over 9 orders of magnitude from subpicosecond to millisecond. The initially locally excited ππ* state rapidly decays within about 100 fs into a charge transfer state following 90° twist motion of the dimethyl amino group. While this twisted intramolecular charge transfer (TICT) state does not trigger any fragmentation, it selectively leads to specific two-color photofragments through absorption of the probe photon at 355 nm. Besides, the optically dark TICT state provides an efficient deactivation path with high intersystem probability to non-dissociative long-lived triplet states. Such a multiscale pump-probe photodissociation scheme paves the way to systematic studies of charge transfer reactions in the excited state of cold ionic systems stored in a cryogenic cooled ion trap and probed continuously up to the millisecond time scale.

Twisted Intramolecular Charge Transfer in Protonated Amino Pyridine.

The excited state properties of protonated ortho (2-), meta (3-), and para (4-) aminopyridine molecules have been investigated through UV photofragmentation spectroscopy and excited state coupled-cluster CC2 calculations. Cryogenic ion spectroscopy allows recording well-resolved vibronic spectroscopy that can be reproduced through Franck-Condon simulations of the ππ* local minimum of the excited state potential energy surface. The excited state lifetimes have also been measured through a pump-probe excitation scheme and compared to the estimated radiative lifetimes. Although protonated aminopyridines are rather simple aromatic molecules, their deactivation mechanisms are indeed quite complex with unexpected results. In protonated 3- and 4-aminopyridine, the fragmentation yield is negligible around the band origin, which implies the absence of internal conversion to the ground state. Besides, a twisted intramolecular charge transfer reaction is evidenced in protonated 4-aminopyridine around the band origin, while excited state proton transfer from the pyridinic nitrogen to the adjacent carbon atom opens with roughly 500 cm(-1) of excess energy.

Non-radiative processes in protonated diazines, pyrimidine bases and an aromatic azine.

The excited state lifetimes of DNA bases are often very short due to very efficient non-radiative processes assigned to the ππ*-nπ* coupling. A set of protonated aromatic diazine molecules (pyridazine, pyrimidine and pyrazine C4H5N2(+)) and protonated pyrimidine DNA bases (cytosine, uracil and thymine), as well as the protonated pyridine (C5H6N(+)), have been investigated. For all these molecules except one tautomer of protonated uracil (enol-keto), electronic spectroscopy exhibits vibrational line broadening. Excited state geometry optimization at the CC2 level has been conducted to find out whether the excited state lifetimes measured from line broadening can be correlated to the calculated ordering of the ππ* and nπ* states and the ππ*-nπ* energy gap. The short lifetimes, observed when one nitrogen atom of the ring is not protonated, can be rationalized by relaxation of the ππ* state to the nπ* state or directly to the electronic ground state through ring puckering.

Trapped Hydronium Radical Produced by Ultraviolet Excitation of Substituted Aromatic Molecule.

The gas phase structure and excited state dynamics of o-aminophenol-H2O complex have been investigated using REMPI, IR-UV hole-burning spectroscopy, and pump-probe experiments with picoseconds laser pulses. The IR-UV spectroscopy indicates that the isomer responsible for the excitation spectrum corresponds to an orientation of the OH bond away from the NH2 group. The water molecule acts as H-bond acceptor of the OH group of the chromophore. The complexation of o-aminophenol with one water molecule induced an enhancement in the excited state lifetime on the band origin. The variation of the excited state lifetime of the complex with the excess energy from 1.4 ± 0.1 ns for the 0-0 band to 0.24 ± 0.3 ns for the band at 0-0 + 120 cm(-1) is very similar to the variation observed in the phenol-NH3 system. This experimental result suggests that the excited state hydrogen transfer reaction is the dominant channel for the non radiative pathway. Indeed, excited state ab initio calculations demonstrate that H transfer leading to the formation of the H3O(•) radical within the complex is the main reactive pathway.

Bonding of heme Fe(III) with dioxygen: observation and characterization of an incipient bond.

While ferrous heme (Fe(II)) within hemoproteins binds dioxygen efficiently, it has not yet been possible to observe the analog complex with ferric heme (Fe(III)). We present the first observation and characterization of the latter complex in a cooled ion trap. The bond formation enthalpy of ferric heme-O2 has been derived from the Van't Hoff equation by means of temperature dependent measurements. The binding energy of the [heme Fe(III)-O2](+) ionic complex is rather strong as compared to that of [heme Fe(III)-N2](+), showing the formation of an incipient Fe-O bond, which is confirmed by the electronic absorption spectra of the two complexes. This first observation of the [heme Fe(III)-O2](+) complex lays the basis for the precise description of its electronic states.

A comprehensive study of cold protonated tyramine: UV photodissociation experiments and ab initio calculations.

We present a comprehensive experimental study of protonated tyramine ions in a cold 3D quadrupole ion trap coupled to a time-of-flight mass spectrometer. Multiple UV photodissociation techniques have been developed, including single and double resonance spectroscopy along with time-resolved excited state lifetime measurements through a picosecond pump-probe scheme. An original UV-UV hole burning method is presented which can be used without modification of the quadrupole ion trap. The electronic spectrum of the cold protonated tyramine exhibits well-defined vibronic transitions, allowing the firm assignment of its two low-lying energy conformations by comparison with CC2 ab initio excited state calculations.