Impact of the graded-gap layer on the admittance of MIS structures based on MBE-grown n-Hg1-xCdxTe (x = 0.22-0.23) with the Al2O3 insulator

The impact of the presence of the near-surface graded-gap layers with an increased content of CdTe on the admittance of MIS structures based on MBE-grown n-Hg1–xCdxTe (x = 0.22–0.23) with the Al2O3 insulating coating has been experimentally studied. It has been shown that the structures with a gradedgap layer are characterized by a deeper and wider capacitance dip in the low-frequency capacitance–voltage (CV) characteristic and by higher values of the differential resistance of the space-charge region than the structures without such a layer. It has been found that the main features of the hysteresis of capacitance dependences typical of the graded-gap structures with SiO2/Si3N4 are also characteristic of the MIS structures with the Al2O3 insulator. The factors that cause an increase in the CV characteristic hysteresis upon formation of the graded-gap layer in structures with SiO2/Si3N4 or Al2O3 are still debatable, although it may be assumed that oxygen plays a certain role in formation of this hysteresis

Admittance of MIS structures based on graded-gap MBE HgCdTe with Al2O3 insulator

The paper presents the results of studies of the admittance of MIS structures based on heteroepitaxial MBE n (p)-Hg0.78Cd0.22Te with insulator coating SiO2/Si3N4 and Al2O3 in the test signal frequency range 10 kHz-1 MHz at temperatures ranging from 8 to 220 K. The main parameters of MIS structures with different insulators were determined. MIS structures with Al2O3 have a large enough insulator capacitance (compared to SiO2/Si3N4), a significant modulation capacitance on the CV characteristics, high dielectric strength and low values of the flat-band voltage. The effective charge density found from the value of the flat-band voltage and slow interface trap density for structures with Al2O3 comparable with the corresponding densities for structures with SiO2/Si3N4

Towards water-free biobanks: long-term dry-preservation at room temperature of desiccation-sensitive enzyme luciferase in air-dried insect cells

© 2017 The Author(s). Desiccation-tolerant cultured cells Pv11 derived from the anhydrobiotic midge embryo endure complete desiccation in an ametabolic state and resume their metabolism after rehydration. These features led us to develop a novel dry preservation technology for enzymes as it was still unclear whether Pv11 cells could preserve an exogenous enzyme in the dry state. This study shows that Pv11 cells protect an exogenous desiccation-sensitive enzyme, luciferase (Luc), preserving the enzymatic activity even after dry storage for 372 days at room temperature. A process including preincubation with trehalose, dehydration, storage, and rehydration allowed Pv11 (Pv11-Luc) cells stably expressing luciferase to survive desiccation and still emit luminescence caused by luciferase after rehydration. Luminescence produced by luciferase in Pv11-Luc cells after rehydration did not significantly decrease in presence of a translation inhibitor, showing that the activity did not derive from de novo enzyme synthesis following the resumption of cell metabolism. These findings indicate that the surviving Pv11 cells almost completely protect luciferase during desiccation. Lacking of the preincubation step resulted in the loss of luciferase activity after rehydration. We showed that preincubation with trehalose associated to induction of desiccation tolerance-related genes in Pv11 cells allowed effective in vivo preservation of enzymes in the dry state

Transcriptome analysis of the anhydrobiotic cell line Pv11 infers the mechanism of desiccation tolerance and recovery

© 2018, The Author(s). The larvae of the African midge, Polypedilum vanderplanki, can enter an ametabolic state called anhydrobiosis to overcome fatal desiccation stress. The Pv11 cell line, derived from P. vanderplanki embryo, shows desiccation tolerance when treated with trehalose before desiccation and resumes proliferation after rehydration. However, the molecular mechanisms of this desiccation tolerance remain unknown. Here, we performed high-throughput CAGE-seq of mRNA and a differentially expressed gene analysis in trehalose-treated, desiccated, and rehydrated Pv11 cells, followed by gene ontology analysis of the identified differentially expressed genes. We detected differentially expressed genes after trehalose treatment involved in various stress responses, detoxification of harmful chemicals, and regulation of oxidoreduction that were upregulated. In the desiccation phase, L-isoaspartyl methyltransferase and heat shock proteins were upregulated and ribosomal proteins were downregulated. Analysis of differentially expressed genes during rehydration supported the notion that homologous recombination, nucleotide excision repair, and non-homologous recombination were involved in the recovery process. This study provides initial insights into the molecular mechanisms underlying the extreme desiccation tolerance of Pv11 cells

Atomic-Level Characterization of the Activation Mechanism of SERCA by Calcium

We have performed molecular dynamics (MD) simulations to elucidate, in atomic detail, the mechanism by which the sarcoplasmic reticulum Ca2+-ATPase (SERCA) is activated by Ca2+. Crystal structures suggest that activation of SERCA occurs when the cytoplasmic head-piece, in an open (E1) conformation stabilized by Ca2+, undergoes a large-scale open-to-closed (E1 to E2) transition that is induced by ATP binding. However, spectroscopic measurements in solution suggest that these structural states (E1 and E2) are not tightly coupled to biochemical states (defined by bound ligands); the closed E2 state predominates even in the absence of ATP, in both the presence and absence of Ca2+. How is this loose coupling consistent with the high efficiency of energy transduction in the Ca2+-ATPase? To provide insight into this question, we performed long (500 ns) all-atom MD simulations starting from the open crystal structure, including a lipid bilayer and water. In both the presence and absence of Ca2+, we observed a large-scale open-to-closed conformational transition within 400 ns, supporting the weak coupling between structural and biochemical states. However, upon closer inspection, it is clear that Ca2+ is necessary and sufficient for SERCA to reach the precise geometrical arrangement necessary for activation of ATP hydrolysis. Contrary to suggestions from crystal structures, but in agreement with solution spectroscopy, the presence of ATP is not required for this activating transition. Principal component analysis showed that Ca2+ reshapes the free energy landscape of SERCA to create a path between the open conformation and the activated closed conformation. Thus the malleability of the free energy landscape is essential for SERCA efficiency, ensuring that ATP hydrolysis is tightly coupled to Ca2+ transport. These results demonstrate the importance of real-time dynamics in the formation of catalytically competent conformations of SERCA, with broad implications for understanding enzymatic catalysis in atomic detail

Towards water-free biobanks: long-term dry-preservation at room temperature of desiccation-sensitive enzyme luciferase in air-dried insect cells

© 2017 The Author(s). Desiccation-tolerant cultured cells Pv11 derived from the anhydrobiotic midge embryo endure complete desiccation in an ametabolic state and resume their metabolism after rehydration. These features led us to develop a novel dry preservation technology for enzymes as it was still unclear whether Pv11 cells could preserve an exogenous enzyme in the dry state. This study shows that Pv11 cells protect an exogenous desiccation-sensitive enzyme, luciferase (Luc), preserving the enzymatic activity even after dry storage for 372 days at room temperature. A process including preincubation with trehalose, dehydration, storage, and rehydration allowed Pv11 (Pv11-Luc) cells stably expressing luciferase to survive desiccation and still emit luminescence caused by luciferase after rehydration. Luminescence produced by luciferase in Pv11-Luc cells after rehydration did not significantly decrease in presence of a translation inhibitor, showing that the activity did not derive from de novo enzyme synthesis following the resumption of cell metabolism. These findings indicate that the surviving Pv11 cells almost completely protect luciferase during desiccation. Lacking of the preincubation step resulted in the loss of luciferase activity after rehydration. We showed that preincubation with trehalose associated to induction of desiccation tolerance-related genes in Pv11 cells allowed effective in vivo preservation of enzymes in the dry state

Transcriptome analysis of the anhydrobiotic cell line Pv11 infers the mechanism of desiccation tolerance and recovery

© 2018, The Author(s). The larvae of the African midge, Polypedilum vanderplanki, can enter an ametabolic state called anhydrobiosis to overcome fatal desiccation stress. The Pv11 cell line, derived from P. vanderplanki embryo, shows desiccation tolerance when treated with trehalose before desiccation and resumes proliferation after rehydration. However, the molecular mechanisms of this desiccation tolerance remain unknown. Here, we performed high-throughput CAGE-seq of mRNA and a differentially expressed gene analysis in trehalose-treated, desiccated, and rehydrated Pv11 cells, followed by gene ontology analysis of the identified differentially expressed genes. We detected differentially expressed genes after trehalose treatment involved in various stress responses, detoxification of harmful chemicals, and regulation of oxidoreduction that were upregulated. In the desiccation phase, L-isoaspartyl methyltransferase and heat shock proteins were upregulated and ribosomal proteins were downregulated. Analysis of differentially expressed genes during rehydration supported the notion that homologous recombination, nucleotide excision repair, and non-homologous recombination were involved in the recovery process. This study provides initial insights into the molecular mechanisms underlying the extreme desiccation tolerance of Pv11 cells

New group of transmembrane proteins associated with desiccation tolerance in the anhydrobiotic midge Polypedilum vanderplanki

© 2020, The Author(s). Larvae of the sleeping chironomid Polypedilum vanderplanki are known for their extraordinary ability to survive complete desiccation in an ametabolic state called “anhydrobiosis”. The unique feature of P. vanderplanki genome is the presence of expanded gene clusters associated with anhydrobiosis. While several such clusters represent orthologues of known genes, there is a distinct set of genes unique for P. vanderplanki. These include Lea-Island-Located (LIL) genes with no known orthologues except two of LEA genes of P. vanderplanki, PvLea1 and PvLea3. However, PvLIL proteins lack typical features of LEA such as the state of intrinsic disorder, hydrophilicity and characteristic LEA_4 motif. They possess four to five transmembrane domains each and we confirmed membrane targeting for three PvLILs. Conserved amino acids in PvLIL are located in transmembrane domains or nearby. PvLEA1 and PvLEA3 proteins are chimeras combining LEA-like parts and transmembrane domains, shared with PvLIL proteins. We have found that PvLil genes are highly upregulated during anhydrobiosis induction both in larvae of P. vanderplanki and P. vanderplanki-derived cultured cell line, Pv11. Thus, PvLil are a new intriguing group of genes that are likely to be associated with anhydrobiosis due to their common origin with some LEA genes and their induction during anhydrobiosis

Admittance of MIS structures based on graded-gap MBE HgCdTe with Al2O3 insulator

The paper presents the results of studies of the admittance of MIS structures based on heteroepitaxial MBE n (p)-Hg0.78Cd0.22Te with insulator coating SiO2/Si3N4 and Al2O3 in the test signal frequency range 10 kHz-1 MHz at temperatures ranging from 8 to 220 K. The main parameters of MIS structures with different insulators were determined. MIS structures with Al2O3 have a large enough insulator capacitance (compared to SiO2/Si3N4), a significant modulation capacitance on the CV characteristics, high dielectric strength and low values of the flat-band voltage. The effective charge density found from the value of the flat-band voltage and slow interface trap density for structures with Al2O3 comparable with the corresponding densities for structures with SiO2/Si3N4

Impact of the graded-gap layer on the admittance of MIS structures based on MBE-grown n-Hg1-xCdxTe (x = 0.22-0.23) with the Al2O3 insulator

The impact of the presence of the near-surface graded-gap layers with an increased content of CdTe on the admittance of MIS structures based on MBE-grown n-Hg1–xCdxTe (x = 0.22–0.23) with the Al2O3 insulating coating has been experimentally studied. It has been shown that the structures with a gradedgap layer are characterized by a deeper and wider capacitance dip in the low-frequency capacitance–voltage (CV) characteristic and by higher values of the differential resistance of the space-charge region than the structures without such a layer. It has been found that the main features of the hysteresis of capacitance dependences typical of the graded-gap structures with SiO2/Si3N4 are also characteristic of the MIS structures with the Al2O3 insulator. The factors that cause an increase in the CV characteristic hysteresis upon formation of the graded-gap layer in structures with SiO2/Si3N4 or Al2O3 are still debatable, although it may be assumed that oxygen plays a certain role in formation of this hysteresis