Thriving at Low pH: Adaptation Mechanisms of Acidophiles

Acid resistance of acidophiles is the result of long-term co-evolution and natural selection of acidophiles and their natural habitats, and formed a relatively optimal acid-resistance network in acidophiles. The acid tolerance network of acidophiles could be classified into active and passive mechanisms. The active mechanisms mainly include the proton efflux and consumption systems, generation of reversed transmembrane electrical potential, and adjustment of cell membrane composition; the passive mechanisms mainly include the DNA and protein repair systems, chemotaxis and cell motility, and quorum sensing system. The maintenance of pH homeostasis is a cell-wide physiological process that adopt differently adjustment strategies, deployment modules, and integration network depending on the cell’s own potential and its habitat environments. However, acidophiles exhibit obvious strategies and modules similarities on acid resistance because of the long-term evolution. Therefore, a comprehensive understanding of acid tolerance network of acidophiles would be helpful for the intelligent manufacturing and industrial application of acidophiles.

Nanoparticles and Nanomaterials as Plant Biostimulants

Biostimulants are materials that when applied in small amounts are capable of promoting plant growth. Nanoparticles (NPs) and nanomaterials (NMs) can be considered as biostimulants since, in specific ranges of concentration, generally in small levels, they increase plant growth. Pristine NPs and NMs have a high density of surface charges capable of unspecific interactions with the surface charges of the cell walls and membranes of plant cells. In the same way, functionalized NPs and NMs, and the NPs and NMs with a corona formed after the exposition to natural fluids such as water, soil solution, or the interior of organisms, present a high density of surface charges that interact with specific charged groups in cell surfaces. The magnitude of the interaction will depend on the materials adhered to the corona, but high-density charges located in a small volume cause an intense interaction capable of disturbing the density of surface charges of cell walls and membranes. The electrostatic disturbance can have an impact on the electrical potentials of the outer and inner surfaces, as well as on the transmembrane electrical potential, modifying the activity of the integral proteins of the membranes. The extension of the cellular response can range from biostimulation to cell death and will depend on the concentration, size, and the characteristics of the corona.

Nanoparticles and Nanomaterials as Plant Biostimulants

Biostimulants are materials that when applied in small amounts are capable of promoting plant growth. Nanoparticles (NPs) and nanomaterials (NMs) can be considered as biostimulants since, in specific ranges of concentration, generally in small levels, they increase the plant growth. Pristine NPs and NMS have a high density of surface charges capable of unspecific interactions with the surface charges of the cell walls and membranes of plant cells. In the same way, the functionalized NPs and NMS, and the NPs and NMs with a corona formed after the exposition to natural fluids such as water, soil solution, or the interior of organisms, presents a high density of surface charges that interact with specific charged groups in cell surfaces. The magnitude of the interaction will depend on the materials adhered to the corona, but the high-density charges located in a small volume causes an intense interaction capable of disturbing the density of surface charges of cell walls and membranes. The electrostatic disturbance can have an impact on the electrical potentials of the outer and inner surfaces, as well as on the transmembrane electrical potential, modifying the activity of the integral proteins of the membranes. The extension of the cellular response can range from biostimulation to cell death and will depend on the concentration, size, and the characteristics of the corona.

Safety Aspect ofEnterococcus faeciumFL31 Strain and Antibacterial Mechanism of Its Hydroxylated Bacteriocin BacFL31 againstListeria monocytogenes

In previous work we have isolated and identified a new strain calledEnterococcus faeciumFL31. The active compound secreted by this strain, “BacFL31”, has been purified and characterized. In the present study, safety aspect, assessed by microbiological and molecular tests, demonstrated thatEnterococcus faeciumFL31 was susceptible to relevant antibiotics, free of hemolytic, gelatinase, DNase, and lipase activities. In addition, it did not harbor virulence and antibiotic resistance genes. Combined SYTOX Green dye and UV-absorbing experiments, along with released extracellular potassium and transmembrane electrical potential measurements, showed that pure BacFL31 at a concentration of 1×MIC (50 μg/mL) could damage cytoplasmic membrane of the pathogenListeria monocytogenesATCC19117. The same concentration causes the leakage of its intracellular constituents and leads to the destruction of this pathogenic microorganism. In summary, this work reflected characteristics ofEnterococcus faeciumFL31 strain and its bacteriocin in terms of functional and safety perspectives.

Structural basis for activation of voltage sensor domains in an ion channel TPC1

Voltage-sensing domains (VSDs) couple changes in transmembrane electrical potential to conformational changes that regulate ion conductance through a central channel. Positively charged amino acids inside each sensor cooperatively respond to changes in voltage. Our previous structure of a TPC1 channel captured an example of a resting-state VSD in an intact ion channel. To generate an activated-state VSD in the same channel we removed the luminal inhibitory Ca2+-binding site (Cai2+), which shifts voltage-dependent opening to more negative voltage and activation at 0 mV. Cryo-EM reveals two coexisting structures of the VSD, an intermediate state 1 that partially closes access to the cytoplasmic side but remains occluded on the luminal side and an intermediate activated state 2 in which the cytoplasmic solvent access to the gating charges closes, while luminal access partially opens. Activation can be thought of as moving a hydrophobic insulating region of the VSD from the external side to an alternate grouping on the internal side. This effectively moves the gating charges from the inside potential to that of the outside. Activation also requires binding of Ca2+ to a cytoplasmic site (Caa2+). An X-ray structure with Caa2+ removed and a near-atomic resolution cryo-EM structure with Cai2+ removed define how dramatic conformational changes in the cytoplasmic domains may communicate with the VSD during activation. Together four structures provide a basis for understanding the voltage-dependent transition from resting to activated state, the tuning of VSD by thermodynamic stability, and this channel’s requirement of cytoplasmic Ca2+ ions for activation.

The structural basis for activation of voltage sensor domains in an ion channel TPC1

Voltage sensing domains (VSDs) couple changes in transmembrane electrical potential to conformational changes that regulate ion conductance through a central channel. Positively charged amino acids inside each sensor cooperatively respond to changes in voltage. Our previous structure of a TPC1 channel captured the first example of a resting-state VSD in an intact ion channel. To generate an activated state VSD in the same channel we removed the luminal inhibitory Ca2+-binding site (Cai2+), that shifts voltage-dependent opening to more negative voltage and activation at 0 mV. Cryo-EM reveals two coexisting structures of the VSD, an intermediate state 1 that partially closes access to the cytoplasmic side, but remains occluded on the luminal side and an intermediate activated state 2 in which the cytoplasmic solvent access to the gating charges closes, while luminal access partially opens. Activation can be thought of as moving a hydrophobic insulating region of the VSD from the external side, to an alternate grouping on the internal side. This effectively moves the gating charges from the inside potential to that of the outside. Activation also requires binding of Ca2+ to a cytoplasmic site (Caa2+). An X-ray structure with Caa2+ removed and a near-atomic resolution cryo-EM structure with Cai2+ removed define how dramatic conformational changes in the cytoplasmic domains may communicate with the VSD during activation. Together four structures provide a basis for understanding the voltage dependent transition from resting to activated state, the tuning of VSD by thermodynamic stability, and this channel’s requirement of cytoplasmic Ca2+-ions for activation.

Talking about bioelectrical potentials using rings of the mesenteric artery without glass micropipettes

In the present study, a practical activity is proposed to adopt an experimental approach to demonstrate the relationship between the equilibrium potential for K+ and transmembrane electrical potential without glass micropipettes. A conventional setup for recording contractile activity of isolated smooth muscle preparations was used based on the events elegantly described by Somlyo and Somlyo in the 1960s. They showed that, in response to a given stimulus, smooth muscle cells may contract, recruiting electromechanical or pharmacomechanical coupling by mechanisms that involve, or not, changes in transmembrane potential, respectively. By means of contractions and relaxations of a ring-like preparation from the rat mesenteric artery, it is possible to observe the functional consequences of handling K+ concentration in the extracellular compartment and the effects caused by opening K+ channels in that preparation, which are significant when the cell membrane establishes an electrical potential difference between intra- and extracellular compartments (driven mainly by K+ permeability under resting conditions). The effects observed by students fit well with values predicted by Nernst and Goldman-Hodgin-Katz equations, and we demonstrated that the activity is able to improve students' comprehension regarding basic principles of bioelectricity.