Adiabatic-passage cross polarization in N-15 NMR spectroscopy of peptides weakly associated to phospholipids: Determination of large RDC

Structural information can be extracted from one-bond residual dipolar couplings (RDC) measured in NMR spectra of systems in field-ordered media. RDC can be on the order of J-couplings if the anisotropy of alignment is ~ 10−2, 10-fold stronger than that typically used for structural studies of water-soluble proteins. In such systems the performance of 1H→ 15N polarization transfer methods of the INEPT type is not satisfactory. In this study we show the effectiveness of adiabatic-passage cross-polarization (APCP) in transferring the 1H→ 15N polarization in the bicelle-associated peptide Leucine Enkephalin (Lenk). APCP is efficient both in static samples and in samples spun at the magic angle (MAS) or any other angle of the spinning axis to the magnetic field (variable-angle spinning, VAS). The anisotropic spectrum of an aligned static sample and the isotropic spectrum of the sample under MAS provide a set of possible values for the 1H-15N RDC of phospholipid-associated Lenk. The unambiguous determination of the 1H-15N RDC was accomplished by means of VAS experiment

Correlation and response relevance in sequence learning

We tested the effects of introducing a secondary sequence into the serial reaction time task. Specifically, we examined the role of correlated streams of information and response relevance. In the first experiment, the order of stimulus locations was correlated with the order of key press responses in the conventional way. A symbol-identity sequence, of a different length, was also present but no manual responses were made to it, and it was not correlated with any other stream of information. In the second experiment, two concurrent streams of location-based stimuli were presented. Both were sequenced but only one sequence required responses. Importantly, the sequences were either correlated with one another or not (same vs. different lengths). In the third experiment, the same design was used but with one sequence visual and the other auditory. In all three experiments, participants became sensitive to the sequence that required responses, and resultant knowledge was largely explicit. They were also sensitive to the sequence that did not require responses but only when it was correlated with the sequence that did, and here resultant knowledge was implicit. The findings suggest that the presence of a secondary sequence can affect learning, but only when stimuli in that sequence are integrated, through correlation, with responses made to the primary sequenc

Incidental task sequence learning: perceptual rather than conceptual?

In four experiments we investigated whether incidental task sequence learning occurs when no instructional task cues are available (i.e. with univalent stimuli). We manipulated task sequence by presenting three simple binary-choice tasks (colour, form or letter case decisions) in regular repeated or random order. Participants were required to use the same two response keys for each of the tasks. We manipulated response sequence by ordering the stimuli so as to produce either a regular or a random order of left versus right-hand key presses. When sequencing in both, or either, separate stream (i.e. task sequence and/or response sequence) was changed to random, only those participants who had processed both sequences together showed evidence of sequence learning in terms of significant response time disruption (Experiments 1-3). This effect disappeared when the sequences were uncorrelated (Experiment 4). The results indicate that only the correlated integration of task sequence and response sequence produced a reliable incidental learning effect. As this effect depends on the predictable ordering of stimulus categories, it suggests that task sequence learning is perceptual rather than conceptual in natur

The bivalency effect: adjustment of cognitive control without response set priming

The occasional occurrence of bivalent stimuli, that is, stimuli with features relevant to two tasks, slows performance on subsequent tasks with univalent stimuli, including those which have no common features with bivalent stimuli (i.e., the "bivalency effect”). We have suggested that the bivalency effect might stem from an episodic context binding arising from the occasional occurrence of bivalent stimuli. However, as the same response set is used usually for univalent and bivalent stimuli, bivalent stimulus features may be negatively primed via response features. We investigated this possibility in two experiments, in which one group of participants used the same response keys for all tasks and another group used separate response keys. The results showed a comparable bivalency effect in both groups. Thus, it rather results from episodic context binding than from response set primin

Semantic representation of synaesthesia

Synaesthesia has multifaceted consequences for both subjective experience and cognitive performance. Here, I broach the issue of how synaesthesia is represented in semantic memory. I hypothesize that, for example, in grapheme colour synaesthesia, colour is represented as an additional feature in the semantic network that enables the formation of associations that are not present in nonsynaesthetes. Thus, synaesthesia provokes richer memory representations which enable learning opportunities that are not present in non-synaesthetes, provides additional memory cues, and may trigger creative ideas

Structural Information from Solid-State NMR

The application of selective averaging techniques combined with multidimensional spectroscopy to solid-state NMR structure determination is discussed. Examples include spin-diffusion and spy-diffusion experiments

NMR of bicelles: orientation and mosaic spread of the liquid-crystal director under sample rotation

Model-membrane systems composed of liquid-crystalline bicellar phases can be uniaxially oriented with respect to a magnetic field, thereby facilitating structural and dynamics studies of membrane-associated proteins. Here we quantitatively characterize a method that allows the manipulation of the direction of this uniaxial orientation. Bicelles formed from DMPC/DHPC are examined by 31P NMR under variable-angle sample-spinning (VAS) conditions, confirming that the orientation of the liquid-crystalline director can be influenced by sample spinning. The director is perpendicular to the rotation axis when Θ (the angle between the sample-spinning axis and the magnetic field direction) is smaller than the magic angle, and is parallel to the rotation axis when Θ is larger than the magic angle. The new 31P NMR VAS data presented are considerably more sensitive to the orientation of the bicelle than earlier 2H studies and the analysis of the sideband pattern allows the determination of the orientation of the liquid-crystal director and its variation over the sample, i.e., the mosaic spread. Under VAS, the mosaic spread is small if Θ deviates significantly from the magic angle but becomes very large at the magic angl

Switched-angle spinning applied to bicelles containing phospholipid-associated peptides

In a model study, the proton NMR spectrum of the opioid pentapeptide leucine-enkephalin associated with bicelles is investigated. The spectral resolution for a static sample is limited due to the large number of anisotropic interactions, in particular strong proton-proton couplings, but resolution is greatly improved by magic-angle sample spinning. Here we present two-dimensional switched-angle spinning NMR experiments, which correlate the high-resolution spectrum of the membrane-bound peptide under magic-angle spinning with its anisotropic spectrum, leading to well-resolved spectra. The two-dimensional spectrum allows the exploitation of the high resolution of the isotropic spectrum, while retaining the structural information imparted by the anisotropic interactions in the static spectrum. Furthermore, switched-angle spinning techniques are demonstrated that allow one to record the proton spectrum of ordered bicellar phases as a function of the angle between the rotor axis and the magnetic field direction, thereby scaling the dipolar interactions by a predefined facto

Solid-state NMR spectroscopy of 10% 13C labeled ubiquitin: spectral simplification and stereospecific assignment of isopropyl groups

We describe the simplification of 13C-13C correlation spectra obtained from a microcrystalline protein sample expressed on a growth medium of 10% fully 13C labeled glucose diluted in 90% natural abundance glucose as compared to a fully labeled sample. Such a labeling scheme facilitates the backbone and side-chain resonance assignment of Phe, Tyr, His, Asp, Asn, Ile, Lys and Pro and yields an unambiguous stereospecific assignment of the valine Cγ1, Cγ2 13C resonances and of Leucine Cδ

Gamifying Intention Memory – Revisiting Ovsiankina

The Ovsiankina effect refers to the desire to complete interrupted and unfinished tasks. In this study, we investigated whether this effect can be generalized and is observable in videogames. We created 16 short videogames, which were comparable in their controls, and half of which were interrupted midway, whilst the other half could be finished. To complete the videogame, participants either had to reach a certain score or achieve the highest score possible in a finite amount of time. After playing, participants chose eight games for replay. Overall, interrupted videogames were chosen more frequently for replay than finished videogames. Thus, we showed that the Ovsiankina effect appears to be robust and observable even in videogames