A unifying perspective of the ultrafast photo-dynamics of Orange Carotenoid Protein from Synechocystis: peril of high-power excitation, existence of different S* states and influence of tagging

A substantial number of Orange Carotenoid Protein (OCP) studies have aimed to describe the evolution of singlet excited states leading to the formation of photo-activated form, OCPR. The most recent one suggests that three picosecond-lived excited states are formed after the sub-100 fs decay of the initial S2 state. The S* state which has the longest reported lifetime of a few to tens of picoseconds is considered to be the precursor of the first red photoproduct P1. Here, we report the ultrafast photo-dynamics of the OCP from Synechocystis PCC 6803, carried out using Visible-NIR femtosecond time-resolved absorption spectroscopy as a function of the excitation pulse power and wavelength. We found that a carotenoid radical cation can form even at relatively low excitation power, obscuring the determination of photo-activation yields for P1. Moreover, the comparison of green (540 nm) and blue (470 nm) excitations revealed the existence of an hitherto uncharacterized excited state, denoted as S~, living a few tens of picoseconds and formed only upon 470 nm excitation. Since neither the P1 quantum yield nor the photo-activation speed over hundreds of seconds vary under green and blue continuous irradiation, this S~ species is unlikely to be involved in the photo-activation mechanism leading to OCPR. We also addressed the effect of His-tagging at the N- or C-termini on excited state photo-physical properties. Differences in spectral signatures and lifetimes of the different excited states were observed, at variance with the usual assumption that His-tagging hardly influences protein dynamics and function. Altogether our results advocate for careful consideration of the excitation power and His-tag position when comparing the photo-activation of different OCP variants, and beg to revisit the notion that S* is the precursor of photoactivated OCPR.

The exchange of the fast substrate water in the S2 state of photosystem II is limited by diffusion of bulk water through channels – implications for the water oxidation mechanism

The molecular oxygen we breathe is produced from water-derived oxygen species bound to the Mn4CaO5 cluster in photosystem II (PSII). Present research points to the central oxo-bridge O5 as the...

Subject matter experts’ feedback on a prototype development of an audio, visual, and haptic phishing email alert system

Phishing emails, also defined as email spam messages, present a threat to both personal and organizational data loss. About 93% of cybersecurity incidents are due to phishing and/or social engineering. Users are continuing to click on phishing links in emails even after phishing awareness training. Thus, it appears that there is a strong need for creative ways to alert and warn users to signs of phishing in emails. ‘System 2 Thinking Mode’ (S2) describes an individual in a more aware state of mind when making important decisions. Ways to trigger S2 include audio alerts, visual alerts, and haptic/vibrations. Assisting the user in noticing signs of phishing in emails could possibly be studied through the delivery of audio, visual, and haptic (vibration) alerts and warnings. This study outlines the empirical results from 32 Subject Matter Experts (SMEs) on an initial prototype design and development of an email phishing alert and warning system. The prototype will be developed to alert and warn users to the signs of phishing in emails in an attempt to switch them to an S2 state of mind. The preliminary results of the SMEs indicated that several features for a phishing alert and warning system could be assembled, resulting in a mobile phishing alert and warning prototype. Visual icons were chosen for each sign of phishing used in the prototype, as well as voice over warnings and haptic vibrations. The preliminary results also determined task measurements, ‘ability to notice’, and ‘time to notice’ signs of phishing in emails.

The geometry relaxation and photodeactivation from the S2 state of dibenzofuran studied by ultrafast spectroscopy

Dibenzofuran (DBF) has attracted much attention from scientists recently since it is applied as a photoluminescence material and pharmaceutically active compound. Since the polychlorinated derivatives are highly toxic and manifest photostability in the environment. Femtosecond transient absorption spectroscopy associated with quantum chemical calculations are employed to investigate the ultrafast excited state dynamics of dibenzofuran from the S2 state in 1,4-dioxane and ethanol, respectively. Following excitation at a wavelength of 266 nm, the S2 state is firstly populated in the Franck–Condon region and preserves the planar molecular structure of the ground state. The observed increase of the transient absorption spectra of the excited state within the first several picoseconds indicates a geometry relaxation occurring on the S2 potential energy surface. The subsequent kinetic traces of excited state absorption show that the S2 state in the adiabatic region decays to the S1 state through a fast internal conversion, followed by intersystem crossing to the T1 state with a decay time of tens and hundreds of picoseconds in ethanol and 1,4-dioxane, respectively. Finally, the deactivation processes from the S1 or T state are slow and take place on a time scale of about 20 ns.

Capturing structural changes of photosystem II using time-resolved serial femtosecond crystallography with proper flash excitations

Photosystem II (PSII) catalyzes light-induced water oxidation through an Si-state cycle, leading to the generation of di-oxygen, protons, and electrons. Pump-probe time-resolved serial femtosecond crystallography (TR-SFX) has been used to capture intermediate states of light-driven enzymatic reactions. In this approach, it is crucial to avoid contamination of light into the samples when analyzing a particular reaction intermediate. Here, we describe a method for determining a proper light condition that avoids light contamination to the PSII microcrystals while minimizing the sample consumption in TR-SFX. With the proper illumination conditions determined, we analyzed the S2-state structure of PSII at room temperature, revealing the structural changes during the S1-to-S2 transition at an ambient temperature. By comparing with the previous studies performed at a low temperature or with a different delay time, we reveal the possible channels for water inlet and proton egress, as well as structural changes important for the water-splitting reaction.