Remodeling of photosynthetic electron transport in Synechocystis sp. PCC 6803 for future hydrogen production from water.

Photosynthetic microorganisms such as the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis) can be exploited for the light-driven synthesis of valuable compounds. Thermodynamically, it is most beneficial to branch-off photosynthetic electrons at ferredoxin (Fd), which provides electrons for a variety of fundamental metabolic pathways in the cell, with the ferredoxin-NADP+ Oxido-Reductase (FNR, PetH) being the main target. In order to re-direct electrons from Fd to another consumer, the high electron transport rate between Fd and FNR has to be reduced. Based on our previous in vitro experiments, corresponding FNR-mutants at position FNR_K190 (Wiegand, K., et al.: "Rational redesign of the ferredoxin-NADP-oxido-reductase/ferredoxin-interaction for photosynthesis-dependent H2-production". Biochim Biophys Acta, 2018) have been generated in Synechocystis cells to study their impact on the cellular metabolism and their potential for a future hydrogen-producing design cell. Out of two promising candidates, mutation FNR_K190D proved to be lethal due to oxidative stress, while FNR_K190A was successfully generated and characterized: The light induced NADPH formation is clearly impaired in this mutant and it shows also major metabolic adaptations like a higher glucose metabolism as evidenced by quantitative mass spectrometric analysis. These results indicate a high potential for the future use of photosynthetic electrons in engineered design cells - for instance for hydrogen production. They also show substantial differences of interacting proteins in an in vitro environment vs. physiological conditions in whole cells.

Cortical networks for auditory detection with and without informational masking: Task effects and implications for conscious perception.

Ambiguous and masked stimuli have been used to study conscious perception by comparing neural activity during different percepts of identical physical stimuli. One limitation of this approach is that it typically requires a reporting task that may engage neural processes beyond those required for conscious perception. Here, we explored potential fMRI correlates of auditory conscious perception with and without overt report. In experiment 1, regular tone patterns were presented as targets under informational masking, and participants reported their percepts on each trial. In experiment 2, regular tone patterns were presented without masking, while the uninformed participants (i) passively fixated, (ii) performed an orthogonal visual task, and (iii) reported trial-wise the presence of the auditory pattern as in experiment 1 (in fixed order). Under informational masking, target-pattern detection was associated with activity in auditory cortex, superior temporal sulcus, and a distributed fronto-parieto-insular network. Unmasked and task-irrelevant tone patterns elicited activity that overlapped with the network observed under informational masking in auditory cortex, the right superior temporal sulcus, and the ventral precentral sulcus in an ROI analysis. We therefore consider these structures candidate regions for a neural substrate of auditory conscious perception. In contrast, activity in the intraparietal sulcus, insula, and dorsal precentral sulcus were only observed for unmasked tone patterns when they were task relevant. These areas therefore appear more closely related to task performance or top-down attention rather than auditory conscious perception, per se.

Cloning, expression, crystallization and preliminary X-ray studies of the ferredoxin-NAD(P)+ reductase from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1.

Ferredoxin-NADP(+) reductase (FNR) is a flavoenzyme that catalyses the reduction of NADP(+) in the final step of the photosynthetic electron-transport chain. FNR from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 (TeFNR) contains an additional 9 kDa domain at its N-terminus relative to chloroplastic FNRs and is more thermostable than those from mesophilic cyanobacteria. With the aim of understanding the structural basis of the thermostability of TeFNR and assigning a structural role to the small additional domain, the gene encoding TeFNR with and without an additional domain was engineered for heterologous expression and the recombinant proteins were purified and crystallized. Crystals of TeFNR without the additional domain belonged to space group P2(1), with unit-cell parameters a = 55.05, b = 71.66, c = 89.73 Å, α = 90, ß = 98.21, γ = 90°.

Correlates of perceptual awareness in human primary auditory cortex revealed by an informational masking experiment.

The presence of an auditory event may remain undetected in crowded environments, even when it is well above the sensory threshold. This effect, commonly known as informational masking, allows for isolating neural activity related to perceptual awareness, by comparing repetitions of the same physical stimulus where the target is either detected or not. Evidence from magnetoencephalography (MEG) suggests that auditory-cortex activity in the latency range 50-250 ms is closely coupled with perceptual awareness. Here, BOLD fMRI and MEG were combined to investigate at which stage in the auditory cortex neural correlates of conscious auditory perception can be observed. Participants were asked to indicate the perception of a regularly repeating target tone, embedded within a random multi-tone masking background. Results revealed widespread activation within the auditory cortex for detected target tones, which was delayed but otherwise similar to the activation of an unmasked control stimulus. The contrast of detected versus undetected targets revealed activity confined to medial Heschl's gyrus, where the primary auditory cortex is located. These results suggest that activity related to conscious perception involves the primary auditory cortex and is not restricted to activity in secondary areas.

Response coding in the Simon task.

Recent findings indicate that two distinct mechanisms can contribute to a Simon effect: a visuomotor information transmission on the one hand and a cognitive code interference on the other hand (see for e.g., Wiegand & Wascher, in Journal of Experimental Psychology: Human Perception and Performance 2005a). Furthermore, it was proposed that the occurrence of one or the other mechanism strongly depends on the way responses are coded. Visuomotor information transmission seems to depend on a correspondence between stimulus position and spatial anatomical status of the effector, whereas cognitive code interference is thought to be based on relative response location codes. To further test the spatial anatomic coding hypothesis, three experiments were conducted, in which the Simon effect with unimanual responses was investigated for horizontal (Experiment 1 and 2) and vertical (Experiment 3) stimulus-response (S-R) relations. Based on the finding of a decreasing effect function (indicating the presence of visuomotor information transmission) for horizontal and vertical S-R relations, it was concluded that visuomotor information transmission occurs whenever there is an overlap between the spatial stimulus feature and parameters of the motor representation of the response. Furthermore, the specific motor representation seems to be task dependent, that is, it entails those response parameters that clearly differentiate between the two response alternatives in a given task situation.

The Simon effect for vertical S-R relations: changing the mechanism by randomly varying the S-R mapping rule?

Recent findings indicate that the shape of the effect function of the Simon effect--derived by analysing the response time distribution for corresponding and non-corresponding trials--differs depending on the task. Specifically, decreasing effect functions have been reported for horizontal and stable rather increasing effect functions have been obtained for vertical stimulus-response (S-R) relations. Furthermore, it has been assumed that these differences reflect distinct mechanisms underlying the Simon effect. However, in two studies decreasing effect functions were reported for the vertical dimension. In order to investigate these contradictory findings four experiments were conducted. Since both studies--in which a decreasing effect function was obtained--used a modified version of the Simon task, that is, randomly varying S-R mapping rules, the Simon effect for fixed and random S-R mapping rules was investigated using vertical (Experiments 1, 2, and 4) and horizontal (Experiment 3) S-R relations. The results indicate that randomly varying S-R mapping rules affect the shape of the effect function for vertical but not for horizontal S-R relations. It was concluded that these findings indicate that the alternation of S-R mapping rules changes the information processing for the vertical dimension in a way, that the same mechanism as for horizontal S-R relations occurs.

Dynamic aspects of stimulus-response correspondence: evidence for two mechanisms involved in the Simon effect.

It has been recently proposed that the time course of the Simon effect may vary across tasks, which might reflect different types of stimulus-response (S-R) transmissions (E. Wascher, U. Schatz, T. Kuder, & R. Verleger, 2001). The authors tested this notion in 4 experiments by comparing Simon effects evoked by horizontal and vertical S-R arrangements. The temporal properties of the effect, as well as lateralized readiness potential-difference waves, indicated a fast and transient influence of the horizontal, but a slow and sustained influence of the vertical spatial stimulus feature on performance. Additional evidence for this temporal dissociation was obtained in experiments that induced a shortening or lengthening of the mean response time. Thus, the data strongly indicate that there are 2 temporally dissociable mechanisms involved in generating the Simon effect for horizontal and vertical S-R relations.