1.
Oscillatory dynamics mechanism induced by protein synthesis time delay in quorum-sensing system.
Chen, M, Liu, H, Yan, F
Physical review. E. 2019;(6-1):062405
Abstract
Recent experimental evidence reports that the oscillatory behavior of quorum sensing plays an extremely important role in the process of bacterial synthesis and release drug to fight cancer. As we know, the six substances AiiA, LuxI, internal AHL, external AHL, AHL substrate, and H_{2}O_{2} are the core parts of the quorum-sensing system. Here, the effects of several important factors, including time delay, variable H_{2}O_{2}, AHL synthesis rate induced by LuxI, and AHL degradation rate induced by AiiA on the oscillatory behavior of the quorum-sensing system are studied theoretically based on a part of mathematical model describing the interaction of the above six substances proposed by Prindle et al. [Nature 508, 387 (2014)10.1038/nature13238]. The results show that the time delay is a prerequisite for inducing oscillation of the quorum-sensing system. Furthermore, the length of time delay can determine the amplitude and period of oscillation. As a further matter, the change of H_{2}O_{2} concentration can induce the oscillatory behavior of the quorum-sensing system. In addition, under the regulation of H_{2}O_{2}, the period robustness of the quorum-sensing system is increased. Similarly, the quorum-sensing system exhibits periodic oscillation when AHL synthesis rate induced by LuxI less than a certain critical value, unless it displays a steady state. Additionally, a too-high or too-low level of AHL degradation rate induced by AiiA will fail to generate oscillation of the quorum-sensing system, only the intermediate level will cause oscillation. Finally, the two and three parameter regions in which the quorum-sensing system exhibits oscillation behavior are generated.
2.
Engineering a Novel Porin OmpGF Via Strand Replacement from Computational Analysis of Sequence Motif.
Lin, M, Zhang, G, Fahie, M, Morgan, LK, Chen, M, Keiderling, TA, Kenney, LJ, Liang, J
Biochimica et biophysica acta. Biomembranes. 2017;(7):1180-1189
Abstract
β-Barrelmembrane proteins (βMPs) form barrel-shaped pores in the outer membrane of Gram-negative bacteria, mitochondria, and chloroplasts. Because of the robustness of their barrel structures, βMPs have great potential as nanosensors for single-molecule detection. However, natural βMPs currently employed have inflexible biophysical properties and are limited in their pore geometry, hindering their applications in sensing molecules of different sizes and properties. Computational engineering has the promise to generate βMPs with desired properties. Here we report a method for engineering novel βMPs based on the discovery of sequence motifs that predominantly interact with the cell membrane and appear in more than 75% of transmembrane strands. By replacing β1-β6 strands of the protein OmpF that lack these motifs with β1-β6 strands of OmpG enriched with these motifs and computational verification of increased stability of its transmembrane section, we engineered a novel βMP called OmpGF. OmpGF is predicted to form a monomer with a stable transmembrane region. Experimental validations showed that OmpGF could refold in vitro with a predominant β-sheet structure, as confirmed by circular dichroism. Evidence of OmpGF membrane insertion was provided by intrinsic tryptophan fluorescence spectroscopy, and its pore-forming property was determined by a dye-leakage assay. Furthermore, single-channel conductance measurements confirmed that OmpGF function as a monomer and exhibits increased conductance than OmpG and OmpF. These results demonstrated that a novel and functional βMP can be successfully engineered through strand replacement based on sequence motif analysis and stability calculation.