scholarly journals Electrophysiological characterization of a diverse group of sugar transporters from Trichoderma reesei

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sami Havukainen ◽  
Jonai Pujol-Giménez ◽  
Mari Valkonen ◽  
Ann Westerholm-Parvinen ◽  
Matthias A. Hediger ◽  
...  
Keyword(s):  
Trichoderma Reesei ◽  
Scientific Literature ◽  
Large Body ◽  
Comprehensive Analysis ◽  
Transport Function ◽  
Sugar Transporters ◽  
New Knowledge ◽  
Electrophysiological Characterization ◽  
Shed Light

AbstractTrichoderma reesei is an ascomycete fungus known for its capability to secrete high amounts of extracellular cellulose- and hemicellulose-degrading enzymes. These enzymes are utilized in the production of second-generation biofuels and T. reesei is a well-established host for their production. Although this species has gained considerable interest in the scientific literature, the sugar transportome of T. reesei remains poorly characterized. Better understanding of the proteins involved in the transport of different sugars could be utilized for engineering better enzyme production strains. In this study we aimed to shed light on this matter by characterizing multiple T. reesei transporters capable of transporting various types of sugars. We used phylogenetics to select transporters for expression in Xenopus laevis oocytes to screen for transport activities. Of the 18 tested transporters, 8 were found to be functional in oocytes. 10 transporters in total were investigated in oocytes and in yeast, and for 3 of them no transport function had been described in literature. This comprehensive analysis provides a large body of new knowledge about T. reesei sugar transporters, and further establishes X. laevis oocytes as a valuable tool for studying fungal sugar transporters.

scholarly journals A databank for intracellular electrophysiological mapping of the adult somatosensory cortex

2018 ◽  
Author(s):  
Angelica da Silva Lantyer ◽  
Niccolò Calcini ◽  
Ate Bijlsma ◽  
Koen Kole ◽  
Melanie Emmelkamp ◽  
...  
Keyword(s):  
Somatosensory Cortex ◽  
Information Transfer ◽  
Intracellular Recordings ◽  
Adult Mice ◽  
Somatic Transformation ◽  
Fast Spiking ◽  
Electrophysiological Characterization ◽  
Neuronal Functions ◽  
Shed Light

AbstractBackgroundNeurons in the supragranular layers of the somatosensory cortex integrate sensory (bottom-up) and cognitive/perceptual (top-down) information as they orchestrate communication across cortical columns. It has been inferred, based on intracellular recordings from juvenile animals, that supragranular neurons are electrically mature by the fourth postnatal week. However, the dynamics of the neuronal integration in the adulthood is largely unknown. Electrophysiological characterization of the active properties of these neurons throughout adulthood will help to address the biophysical and computational principles of the neuronal integration.FindingsHere we provide a database of whole-cell intracellular recordings from 315 neurons located in the supragranular layers (L2/3) of the primary somatosensory cortex in adult mice (9-45 weeks old) from both sexes (females, N=195; males, N=120). Data include 361 somatic current-clamp (CC) and 476 voltage-clamp (VC) experiments, recorded using a step-and-hold protocol (CC, N=257; VC, N=46), frozen noise injections (CC, N=104) and triangular voltage sweeps (VC, 10 (N=132), 50 (N=146) and 100 ms (N=152)), from regular spiking (N=169) and fast-spiking neurons (N=66).ConclusionsThe data can be used to systematically study the properties of somatic integration, and the principles of action potential generation across sexes and across electrically characterized neuronal classes in adulthood. Understanding the principles of the somatic transformation of postsynaptic potentials into action potentials will shed light onto the computational principles of intracellular information transfer in single neurons and information processing in neuronal networks, helping to recreate neuronal functions in artificial systems.

Electrophysiological Characterization of Transport Across Outer Membrane Channels from Gram-Negative Bacteria in Their Native Environment

2019 ◽  
Author(s):  
Jiajun Wang ◽  
Rémi Terrasse ◽  
Jayesh Arun Bafna ◽  
Lorraine Benier ◽  
Mathias Winterhalter
Keyword(s):  
Outer Membrane ◽  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Multi Drug Resistance ◽  
Gram Negative Bacteria ◽  
Membrane Channels ◽  
Gram Negative ◽  
Electrophysiological Characterization

Multi-drug resistance in Gram-negative bacteria is often associated with low permeability of the outer membrane. To investigate the role of membrane channels in the uptake of antibiotics, we extract, purify and reconstitute them into artificial planar membranes. To avoid this time-consuming procedure, here we show a robust approach using fusion of native outer membrane vesicles (OMV) into planar lipid bilayer which moreover allows also to some extend the characterization of membrane protein channels in their native environment. Two major membrane channels from <i>Escherichia coli</i>, OmpF and OmpC, were overexpressed from the host and the corresponding OMVs were collected. Each OMV fusion revealed surprisingly single or only few channel activities. The asymmetry of the OMV´s translates after fusion into the lipid membrane with the LPS dominantly present at the side of OMV addition. Compared to conventional reconstitution methods, the channels fused from OMVs containing LPS have similar conductance but a much broader distribution. The addition of Enrofloxacin on the LPS side yields somewhat higher association (<i>k<sub>on</sub></i>) and lower dissociation (<i>k<sub>off</sub></i>) rates compared to LPS-free reconstitution. We conclude that using outer membrane vesicles is a fast and easy approach for functional and structural studies of membrane channels in the native membrane.

scholarly journals Ni Oxidation State and Ligand Saturation Impact on the Capability of Octaazamacrocyclic Complexes to Bind and Reduce CO2

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4139
Author(s):  
Barbora Vénosová ◽  
Ingrid Jelemenská ◽  
Jozef Kožíšek ◽  
Peter Rapta ◽  
Michal Zalibera ◽  
...  
Keyword(s):  
Quantum Theory ◽  
Oxidation State ◽  
Central Atom ◽  
Population Analysis ◽  
Mulliken Population Analysis ◽  
Mulliken Population ◽  
Theoretical Methods ◽  
Ni Oxidation ◽  
Shed Light

Two 15-membered octaazamacrocyclic nickel(II) complexes are investigated by theoretical methods to shed light on their affinity forwards binding and reducing CO2. In the first complex 1[NiIIL]0, the octaazamacrocyclic ligand is grossly unsaturated (π-conjugated), while in the second 1[NiIILH]2+ one, the macrocycle is saturated with hydrogens. One and two-electron reductions are described using Mulliken population analysis, quantum theory of atoms in molecules, localized orbitals, and domain averaged fermi holes, including the characterization of the Ni-CCO2 bond and the oxidation state of the central Ni atom. It was found that in the [NiLH] complex, the central atom is reduced to Ni0 and/or NiI and is thus able to bind CO2 via a single σ bond. In addition, the two-electron reduced 3[NiL]2− species also shows an affinity forwards CO2.

scholarly journals The Mutant Phenotype Associated With P-Element Alleles of the vestigial Locus in Drosophila melanogaster May Be Caused by a Readthrough Transcript Initiated at the P-Element Promoter

Genetics ◽  
2001 ◽  
Vol 157 (4) ◽  
pp. 1665-1672 ◽  
Author(s):  
Ross B Hodgetts ◽  
Sandra L O'Keefe
Keyword(s):  
Rna Secondary Structure ◽  
Insertion Site ◽  
Large Body ◽  
P Element ◽  
Wild Type ◽  
Subtle Difference ◽  
Pronounced Difference ◽  
Internal Repeat ◽  
Mutant Phenotypes

Abstract We report here the isolation of a new P-element-induced allele of the vestigial locus vg2a33, the molecular characterization of which allows us to propose a unifying explanation of the phenotypes of the large number of vestigial P-element alleles that now exists. The first P-element allele of vestigial to be isolated was vg21, which results in a very weak mutant wing phenotype that is suppressed in the P cytotype. By destabilizing vg2a33 in a dysgenic cross, we isolated the vg2a33 allele, which exhibits a moderate mutant wing phenotype and is not suppressed by the P cytotype. The new allele is characterized by a 46-bp deletion that removes the 3′-proximal copy of the 11-bp internal repeat from the P element of vg21. To understand how this subtle difference between the two alleles leads to a rather pronounced difference in their phenotypes, we mapped both the vg and P-element transcription units present in wild type and mutants. Using both 5′-RACE and S1 protection, we found that P-element transcription is initiated 19 bp farther upstream than previously thought. Using primer extension, the start of vg transcription was determined to lie 435 bp upstream of the longest cDNA recovered to date and upstream of the P-element insertion site. Our discovery that the P element is situated within the first vg exon has prompted a reassessment of the large body of genetic data on a series of alleles derived from vg21. Our current hypothesis to explain the degree of variation in the mutant phenotypes and their response to the P repressor invokes a critical RNA secondary structure in the vg transcript, the formation of which is hindered by a readthrough transcript initiated at the P-element promoter.

scholarly journals The Underlying Order Induced by Orthogonality and the Quantum Speed Limit

2021 ◽  
Vol 3 (3) ◽  
pp. 376-388
Author(s):  
Francisco J. Sevilla ◽  
Andrea Valdés-Hernández ◽  
Alan J. Barrios
Keyword(s):  
Energy Spectrum ◽  
Energy Distribution ◽  
Finite Time ◽  
Complete Characterization ◽  
Comprehensive Analysis ◽  
Orthogonality Condition ◽  
Speed Limit ◽  
Physical Quantities

We perform a comprehensive analysis of the set of parameters {ri} that provide the energy distribution of pure qutrits that evolve towards a distinguishable state at a finite time τ, when evolving under an arbitrary and time-independent Hamiltonian. The orthogonality condition is exactly solved, revealing a non-trivial interrelation between τ and the energy spectrum and allowing the classification of {ri} into families organized in a 2-simplex, δ2. Furthermore, the states determined by {ri} are likewise analyzed according to their quantum-speed limit. Namely, we construct a map that distinguishes those ris in δ2 correspondent to states whose orthogonality time is limited by the Mandelstam–Tamm bound from those restricted by the Margolus–Levitin one. Our results offer a complete characterization of the physical quantities that become relevant in both the preparation and study of the dynamics of three-level states evolving towards orthogonality.

scholarly journals Past and future trends of Egypt’s water consumption and its sources

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Catherine A. Nikiel ◽  
Elfatih A. B. Eltahir
Keyword(s):  
Economic Growth ◽  
Virtual Water ◽  
Comprehensive Analysis ◽  
Future Trends ◽  
Nile Water ◽  
The Gift ◽  
Per Capita ◽  
The 1960S

AbstractFor millennia the Nile supplied Egypt with more water than needed. As the population grew and the economy expanded, demand on water increased accordingly. Here, we present a comprehensive analysis to reconstruct how total demand on water outstripped supply of the Nile water in the late 1970s, starting from a surplus of about 20 km3 per year in the 1960s leading to a deficit of about 40 km3 per year by the late 2010s. The gap is satisfied by import of virtual water. The role of economic growth in driving per capita demand on water is quantified based on detailed analysis of water use by agriculture and other sectors. We develop and test an empirical model of water demand in Egypt that relates demand on water to growth rates in the economy and population. Looking forward, we project that within this decade of the 2020 s, under nominal scenarios of population and economic growth, Egypt is likely to import more virtual water than the water supplied by the Nile, bringing into question the historical characterization of Egypt as “the gift of the Nile”.

scholarly journals Electrophysiological characterization of contact sites in brain mitochondria.

1990 ◽  
Vol 265 (2) ◽  
pp. 908-913
Author(s):  
O Moran ◽  
G Sandri ◽  
E Panfili ◽  
W Stühmer ◽  
M C Sorgato
Keyword(s):  
Brain Mitochondria ◽  
Contact Sites ◽  
Electrophysiological Characterization
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