Photoelectron Circular Dichroism of Aqueous-Phase Alanine

TL;DR

The paper addresses how chirality manifests in aqueous environments by measuring core-level PECD in aqueous-phase alanine using liquid-jet photoelectron spectroscopy with circularly polarized light. The authors report a significant, site-specific PECD for the C1 carbon of alanine, with the signal strongest in the anionic form and showing limited dependence on photoelectron kinetic energy within the studied range. C2 and C3 carbons do not exhibit convincingly resolvable PECD under the current sensitivity, highlighting pronounced site-dependence of PECD in solution. The results demonstrate the feasibility of liquid-phase PECD for biologically relevant molecules, emphasize the influence of solvation and protonation on chiral electronic structure, and outline theoretical and instrumental advances needed to fully interpret solvated PECD phenomena.

Abstract

Amino acids and other small chiral molecules play key roles in biochemistry. However, in order to understand how these molecules behave in vivo, it is necessary to study them under aqueous-phase conditions. Photoelectron circular dichroism (PECD) has emerged as an extremely sensitive probe of chiral molecules, but its suitability for application to aqueous solutions had not yet been proven. Here, we report on our PECD measurements of aqueous-phase alanine, the simplest chiral amino acid. We demonstrate that the PECD response of alanine in water is different for each of alanine's carbon atoms, and is sensitive to molecular structure changes (protonation states) related to the solution pH. For C~1s photoionization of alanine's carboxylic acid group, we report PECD of comparable magnitude to that observed in valence-band photoelectron spectroscopy of gas-phase alanine. We identify key differences between PECD experiments from liquids and gases, discuss how PECD may provide information regarding solution-specific phenomena -- for example the nature and chirality of the solvation shell surrounding chiral molecules in water -- and highlight liquid-phase PECD as a powerful new tool for the study of aqueous-phase chiral molecules of biological relevance.

Figures (4)

• Figure 1: Representative as-measured C 1s photoelectron spectra corresponding to aqueous-phase alanine's three chemically distinct carbons measured with $h\nu$ = 480.46 eV photons under acidic (top), neutral (middle), and basic (bottom) aqueous conditions. Measurements were conducted with 1 M aqueous solutions of L-alanine. The dashed black curves are fits to the data following the subtraction of background signals (dashed purple lines) using exponentially modified Gaussian profiles grushka_exmGauss_1972. The asymmetry parameter was constrained to be between 0.2 and 0.3, and these values were kept constant in fits across different pH conditions. The ratios of peak widths were also constrained to differ by no more than 5% across the three fits. Molecular structures of the dominant protonation state of alanine at the various pH values investigated are shown as insets.
• Figure 2: Top: Illustrative aqueous-phase (pH 13) alanine C 1s photoelectron spectra corresponding to right- and left-handed circularly polarized light (red and blue lines, respectively). The high-energy spectra (right) exhibit low background signal, but vanishing photoelectron circular dichroism (PECD). The low-energy spectra sit atop a large background signal, but reveal significant PECD following subtraction of background signal (purple lines). Bottom: Hypothetical $b^{+1}_{1}$ parameters for each of alanine's carbon groups as a function of peak kinetic energy. The blue shaded zone represents the onset of significant kinetic-energy dependent elastic or quasi-elastic electron scattering. Within the green region, scattering of primary photoelectrons is sufficient to make resolution of the photoelectron features unfeasible. Values of $b^{+1}_{1}$ are currently inaccessible for this system within this kinetic-energy range. Dashed gray boxes represent approximate peak positions for the spectra shown above. The carbon atoms C$_{1}$, C$_{2}$, and C$_{3}$ are identified in Fig. \ref{['Alanine_PECD_clean:fig:XPS']}.
• Figure 3: Top: Representative intensity-scaled C 1s photoelectron spectra of 1 M L-alanine aqueous solution (pH 13) measured at $h\nu=305$ eV for left- and right-handed circularly polarized light (blue and red curves, respectively), along with the calculated percent difference between the spectra for C$_{1}$ group to background subtraction (right axis). Middle: Background-subtracted data, along with calculated percent difference between the spectra for C$_{1}$ (right axis). Bottom: Values of $b^{+1}_{1}$ obtained through the same process for D-, L-, and racemic DL-alanine (red, blue, and green points, respectively). The error bars of these points represent the standard deviation of the percent-difference data shown in the middle panel. The spectra shown here are averages of ten acquisitions, as typical for the data discussed in this study.
• Figure 4: Values of the $b_{1}^{+1}$ photoionization parameter obtained for C 1s measurements of aqueous solutions of D-, L-, and DL-alanine (red, blue, and green points, respectively) at pH 1, 7, and 13 (top, middle, and bottom; corresponding to the cationic, zwitterionic, and anionic form of the molecule, respectively). All $b_{1}^{+1}$ values shown correspond to photoionization of the C$_{1}$ carboxylic group. The data displayed is the result of binning the results of individual measurement sets using a kinetic-energy window of 250 meV. The shaded error bars represent the combined error of the binned data points. A description of the error propagation may be found in the supplementary information.