Elongated Bacterial Pili as a Versatile Alignment Medium for NMR Spectroscopy.

In NMR spectroscopy, residual dipolar couplings (RDCs) have emerged as one of the most exquisite probes of biological structure and dynamics. The measurement of RDCs relies on the partial alignment of the molecule of interest, for example by using a liquid crystal as a solvent. Here, we establish bacterial type 1 pili as an alternative liquid-crystalline alignment medium for the measurement of RDCs. To achieve alignment at pilus concentrations that allow for efficient NMR sample preparation, we elongated wild-type pili by recombinant overproduction of the main structural pilus subunit. Building on the extraordinary stability of type 1 pili against spontaneous dissociation and unfolding, we show that the medium is compatible with challenging experimental conditions such as high temperature, the presence of detergents, organic solvents or very acidic pH, setting it apart from most established alignment media. Using human ubiquitin, HIV-1 TAR RNA and camphor as spectroscopic probes, we demonstrate the applicability of the medium for the determination of RDCs of proteins, nucleic acids and small molecules. Our results show that type 1 pili represent a very useful alternative to existing alignment media and may readily assist the characterization of molecular structure and dynamics by NMR.

Variability in the Vestibulo-Ocular Reflex and Vestibular Perception.

The vestibular system enables humans to estimate self-motion, stabilize gaze and maintain posture, but these behaviors are impacted by neural noise at all levels of processing (e.g., sensory, central, motor). Despite its essential importance, the behavioral impact of noise in human vestibular pathways is not completely understood. Here, we characterize the vestibular imprecision that results from neural noise by measuring trial-to-trial vestibulo-ocular reflex (VOR) variability and perceptual just-noticeable differences (JNDs) in the same human subjects as a function of stimulus intensity. We used head-centered yaw rotations about an Earth-vertical axis over a broad range of motion velocities (0-65°/s for VOR variability and 3-90°/s peak velocity for JNDs). We found that VOR variability increased from approximately 0.6°/s at a chair velocity of 1°/s to approximately 3°/s at 65°/s; it exhibited a stimulus-independent range below roughly 1°/s. Perceptual imprecision ("sigma") increased from 0.76°/s at 3°/s to 4.7°/s at 90°/s. Using stimuli that manipulated the relationship between velocity, displacement and acceleration, we found that velocity was the salient cue for VOR variability for our motion stimuli. VOR and perceptual imprecision both increased with stimulus intensity and were broadly similar over a range of stimulus velocities, consistent with a common noise source that affects motor and perceptual pathways. This contrasts with differing perceptual and motor stimulus-dependent imprecision in visual studies. Either stimulus-dependent noise or non-linear signal processing could explain our results, but we argue that afferent non-linearities alone are unlikely to be the source of the observed behavioral stimulus-dependent imprecision.