A new class of protein sensor links spirochete pleomorphism, persistence, and chemotaxis

Pathogenic spirochetes can alter their morphologies and behaviors to infect and survive within their hosts. Previous reports demonstrate that the formation of so-called round bodies and biofilms, and chemotaxis are involved in spirochete pathogenesis. Here, in the spirochete Treponema denticola, we report a direct link between these cellular states that involves a new class of protein sensor (CheWS) with hitherto unclear function. Using cryo-EM methods, protein modeling, bioinformatics, genetics methods, and behavioral assays we demonstrate that spirochetes regulate these behaviors in response to the small molecule s-adenosylmethionine (SAM) via a SAM sensor that is anchored to chemotaxis arrays. CheWS influences chemotaxis, biofilm and round body formation under non-stressed conditions by a novel sporulation-like mechanism. Taken together, we establish an improved model for round body formation, we discovered a direct link between this SAM sensor and changes in cellular states, as well as characterized a new sensor class involved in chemotaxis.

Development of a Textile Based Protein Sensor for Monitoring the Healing Progress of a Wound

This article focuses on the design and fabrication of flexible textile-based protein sensors to be embedded in wound dressings. Chronic wounds require continuous monitoring to prevent further complications and to determine the best course of treatment in the case of infection. As proteins are essential for the progression of wound healing, they can be used as an indicator of wound status. Through measuring protein concentrations, the sensor can assess and monitor the wound condition continuously as a function of time. The protein sensor consists of electrodes that are directly screen printed using both silver and carbon composite inks on polyester nonwoven fabric which was deliberately selected as this is one of the common backing fabrics currently used in wound dressings. Three sensor designs were investigated to determine if any were suitable for protein detection. These sensors were experimentally evaluated and compared to each other by using albumin protein in phosphate buffered saline (PBS). A comprehensive set of cyclic voltammetry measurements were used to determine the optimal sensor design to provide the measurement of protein in solution. The best sensor was comprised of only silver conductive ink present to form the tracks outside the interface zone and a carbon only layer in the working and counter electrodes at the interface zone. This design prevents the formation of silver dioxide and protects the sensor from rapid decay, which allows for the recording of consecutive measurements using the same sensor. The chosen printed protein sensor was able to detect BSA at varying concentrations ranging from 30-0.3 mg/ml with a sensitivity of 0.0026µA/M.

C-Reactive Protein Sensor Based on Porous Silicon Bragg Stack Interferometers

Bio-functionalized nanomaterials represent the cutting-edge research for a sensing biomolecules in nano-systems. Their physicochemical properties of porous silicon bring along advantages in sensing applications. Here, a modified biosensor based on the anti-C-reactive protein-modified porous silicon Bragg stack interferometer was developed to detect C-reactive protein. The SEM images of the surface and cross-sectional views of the Bragg stack porous silicon exhibited the pore sizes in the 10–20 nm range. The fabrication, optical characterization, and surface derivatization of the interferometer were also reported. This sensor was assessed by measuring the reflection peaks in the white light reflection spectrum. As a result, molecular binding was detected as a shift in the wavelength of these reflection peaks. In addition, a dramatic decrease in the reflectivity was observed in the reflectivity spectrum within 10 s, thereby indicating a C-reactive protein detection limit of 100 pM.

Nanopore dwell time analysis permits sequencing and conformational assignment of pseudouridine in SARS-CoV-2

Nanopore devices can directly sequence RNA, and the method has the potential to determine locations of epitranscriptomic modifications that have grown in significance because of their roles in cell regulation and stress response. Pseudouridine (Ψ), the most common modification in RNA, was sequenced with a nanopore system using a protein sensor with a helicase brake in synthetic RNAs with 100% modification at 18 known human pseudouridinylation sites. The new signals were compared to native uridine (U) control strands to characterize base calling and associated errors as well as ion current and dwell time changes. The data point to strong sequence context effects in which Ψ can easily be detected in some contexts while in others Ψ yields signals similar to U that would be false negatives in an unknown sample. We identified that the passage of Ψ through the helicase brake slowed the translocation kinetics compared to U and showed a smaller sequence bias that could permit detection of this modification in RNA. The unique signals from Ψ relative to U are proposed to reflect the syn-anti conformational flexibility of Ψ not found in U, and the difference in π stacking between these bases. This observation permitted analysis of SARS-CoV-2 nanopore sequencing data to identify five conserved Ψ sites on the 3′ end of the viral sub-genomic RNAs, and other less conserved Ψ sites. Using the helicase as a sensor protein in nanopore sequencing experiments enables detection of this modification in a greater number of relevant sequence contexts. The data are discussed concerning their analytical and biological significance.

A Multichannel Pattern-Recognition-Based Protein Sensor with a Fluorophore-Conjugated Single-Stranded DNA Set

Recently, pattern-recognition-based protein sensing has received considerable attention because it offers unique opportunities that complement more conventional antibody-based detection methods. Here, we report a multichannel pattern-recognition-based sensor using a set of fluorophore-conjugated single-stranded DNAs (ssDNAs), which can detect various proteins. Three different fluorophore-conjugated ssDNAs were placed into a single microplate well together with a target protein, and the generated optical response pattern that corresponds to each environment-sensitive fluorophore was read via multiple detection channels. Multivariate analysis of the resulting optical response patterns allowed an accurate detection of eight different proteases, indicating that fluorescence signal acquisition from a single compartment containing a mixture of ssDNAs is an effective strategy for the characterization of the target proteins. Additionally, the sensor could identify proteins, which are potential targets for disease diagnosis, in a protease and inhibitor mixture of different composition ratios. As our sensor benefits from simple construction and measurement procedures, and uses accessible materials, it offers a rapid and simple platform for the detection of proteins.