Residue-specific N-terminal glycine to aldehyde transformation renders analytically pure single-site labeled proteins.

Here, we present N-Gly-specific glyoxamide generation in native proteins, isolated or in a complex mixture. The resulting aldehyde enables parallel installation of probes and a purification platform to render analytically pure single-site tagged proteins. It renders N-Gly engineered insulin without perturbing its structure, receptor binding, and downstream signaling pathway.

Impact of Genetic Variation in Gene Regulatory Sequences: A Population Genomics Perspective.

The unprecedented rise of high-throughput sequencing and assay technologies has provided a detailed insight into the non-coding sequences and their potential role as gene expression regulators. These regulatory non-coding sequences are also referred to as cis-regulatory elements (CREs). Genetic variants occurring within CREs have been shown to be associated with altered gene expression and phenotypic changes. Such variants are known to occur spontaneously and ultimately get fixed, due to selection and genetic drift, in natural populations and, in some cases, pave the way for speciation. Hence, the study of genetic variation at CREs has improved our overall understanding of the processes of local adaptation and evolution. Recent advances in high-throughput sequencing and better annotations of CREs have enabled the evaluation of the impact of such variation on gene expression, phenotypic alteration and fitness. Here, we review recent research on the evolution of CREs and concentrate on studies that have investigated genetic variation occurring in these regulatory sequences within the context of population genetics.

Efficient correction of armband rotation for myoelectric-based gesture control interface.

OBJECTIVE: The appearance of commercial myoelectric armbands has greatly increased the portability and convenience of myoelectric controlled interfaces (MCIs). However, one limitation of the current state-of-the-art myoelectric control algorithms is that they have poor robustness against armband displacements, especially rotation, leading to great algorithmic performance degradation. The traditional remedy, retraining the interface, requires the data collection of all gestures and is impractical in many applications. The recently proposed position verification (PV) framework focused on quickly identifying and correcting the electrode positions after the displacement, showing the potential to restore the performance of MCI in a faster way. However, its online effectiveness is still yet to be validated. APPROACH: This work proposed a novel algorithm of identifying the rotation direction to improve the efficiency of the PV framework and demonstrated the real-time capability of the PV framework using a commercially available armband. MAIN RESULTS: The results showed that with PV, a 1.5-cm rotation could be corrected with an average of 3.1 ± 1.5 interactive adjustments, equivalent to around 15.5 ± 7.5 s, which was greatly reduced compared to retraining. There was no significant difference in the real-time control performance between before the armband displacement and after the PV correction. SIGNIFICANCE: To the best of our knowledge, this study was the first maintaining pattern recognition-based myoelectric control performance in the presence of electrode shifts without recollecting the entire training data. It suggested the feasibility of the PV framework used in the myoelectric armband and MCI for practical applications.

Exploration of Free Energy Surfaces Across a Membrane Channel Using Metadynamics and Umbrella Sampling.

To reach their site of action, it is essential for antibiotic molecules to cross the bacterial outer membrane. The progress of enhanced sampling techniques in molecular dynamics simulations enables us to understand these translocations at an atomic level. To this end, calculations of free energy surfaces for these permeation processes are of key importance. Herein, we investigate the translocation of a variety of anionic solutes through the outer membrane pore OprO of the Gram-negative bacterium Pseudomonas aeruginosa using the metadynamics and umbrella sampling techniques at the all-atom level. Free energy calculations have been performed employing these two distinct methods in order to illustrate the difference in computed free energies, if any. The investigated solutes range from a single atomic chloride ion over a multiatomic monophosphate ion to a more bulky fosmidomycin antibiotic. The role of complexity of the permeating solutes in estimating accurate free energy profiles is demonstrated by performing extensive convergence analysis. For simple monatomic ions, good agreement between the well-tempered metadynamics and the umbrella sampling approaches is achieved, while for the permeation of the monophosphate ion differences start to appear. In the case of larger molecules such as fosmidomycin it is a tough challenge to achieve converged free energy profiles. This issue is mainly due to neglecting orthogonal degrees of freedom during the free energy calculations. Nevertheless, the freely driven metadynamics approach leads to clearly advantageous results. Additionally, atomistic insights of the translocation mechanisms of all three solutes are discussed.