Crystal Structure Analysis and the Identification of Distinctive Functional Regions of the Protein Elicitor Mohrip2

The protein elicitor MoHrip2, which was extracted from Magnaporthe oryzae as an exocrine protein, triggers the tobacco immune system and enhances blast resistance in rice. However, the detailed mechanisms by which MoHrip2 acts as an elicitor remain unclear. Here, we investigated the structure of MoHrip2 to elucidate its functions based on molecular structure. The 3-dimensional structure of MoHrip2 was obtained. Overall, the crystal structure formed a β-barrel structure and showed high similarity to the pathogenesis-related (PR) thaumatin superfamily protein thaumatin-like xylanase inhibitor (TL-XI). To investigate the functional regions responsible for MoHrip2 elicitor activities, the full length and 8 truncated proteins were expressed in Escherichia coli and were evaluated for elicitor activity in tobacco. Biological function analysis showed that MoHrip2 triggered the defense system against Botrytis cinerea in tobacco. Moreover, only MoHrip2M14 and other fragments containing the 14 amino acids residues in the middle region of the protein showed the elicitor activity of inducing a hypersensitive response and resistance related pathways, which were similar to that of full-length MoHrip2. These results revealed that the central 14 amino acid residues were essential for anti-pathogenic activity

Monolayer MoS2-Based Flexible and Highly Sensitive Pressure Sensor with Wide Sensing Range

Flexible pressure sensors play an important role in flexible robotics, human-machine interaction (HMI), and human physiological information. However, most of the reported flexible pressure sensors suffer from a highly nonlinear response and a significant decrease in sensitivity at high pressures. Herein, we propose a flexible novel iontronic pressure sensor based on monolayer molybdenum disulfide (MoS2). Based on the unique structure and the excellent mechanical properties as well as the large intercalation capacitance of MoS2, the prepared sensor holds an ultra-high sensitivity (Smax = 89.75 kPa−1) and a wide sensing range (722.2 kPa). Further, the response time and relaxation time of the flexible sensor are only 3 ms, respectively, indicating that the device can respond to external pressure rapidly. In addition, it shows long-term cycling stability (over 5000 cycles with almost no degradation) at a high pressure of 138.9 kPa. Finally, it is demonstrated that the sensor can be used in physiological information monitoring and flexible robotics. It is anticipated that our prepared sensor provide a reliable approach to advance the theory and practicality of the flexible sensor electronics

Highly Selective Off–On Fluorescent Probe for Imaging Thioredoxin Reductase in Living Cells

The first fluorescent probe for mammalian thioredoxin reductase (TrxR), TRFS-green, was designed, synthesized, and fully evaluated. The probe features a 1,2-dithiolane scaffold with a quenched naphthalimide fluorophore. TRFS-green displays a green fluorescence off–on change induced by the TrxR-mediated disulfide cleavage and subsequent intramolecular cyclization to liberate the masked naphthalimide fluorophore. It was demonstrated in vitro that TRFS-green manifests high selectivity toward TrxR over other related enzymes and various small molecule thiols as well as biological reducing molecules. HPLC analyses indicated that TRFS-green was exclusively converted to naphthalimide catalyzed by TrxR. The ability in triggering on the fluorescence signal by cellular protein extracts correlates well with the endogenous TrxR activity in different cells. Furthermore, inhibition of TrxR by 2,4-dinitrochlorobenzene or depletion of TrxR by immunoprecipitation remarkably decreases the reduction of TRFS-green by cellular protein extracts. Finally, TRFS-green was successfully applied in imaging TrxR activity in living cells. The fluorescence signal of TRFS-green in living cells was inhibited by pretreating the cells with TrxR inhibitor in a dose-dependent manner, potentiating the development of living cell-based screening assay for identifying TrxR inhibitors. We expect the novel fluorescent probe TRFS-green would facilitate the discovery of TrxR-targeting small molecules for potential therapeutic agents and provide significant advances in understanding the physiological/pathophysiological functions of TrxR in vivo