scholarly journals Electrophysiological characterization of Af-Amt1 on a solid supported membrane

2012 ◽  
Vol 1817 ◽  
pp. S38-S39
Author(s):  
Tobias Wacker ◽  
Juan J. Garcia-Celma ◽  
Philipp Lewe ◽  
Susana L.A. Andrade
Keyword(s):  
Supported Membrane ◽  
Electrophysiological Characterization

scholarly journals Electrophysiological characterization of the archaeal transporter NCX_Mj using solid supported membrane technology

2016 ◽  
Vol 147 (6) ◽  
pp. 485-496 ◽  
Author(s):  
Maria Barthmes ◽  
Jun Liao ◽  
Youxing Jiang ◽  
Andrea Brüggemann ◽  
Christian Wahl-Schott
Keyword(s):  
Functional Data ◽  
Membrane Transporters ◽  
Transporter Proteins ◽  
Supported Membrane ◽  
Functional Studies ◽  
Sodium Calcium ◽  
Calcium Exchange ◽  
Electrophysiological Characterization ◽  
Insight Into

Sodium–calcium exchangers (NCXs) are membrane transporters that play an important role in Ca2+ homeostasis and Ca2+ signaling. The recent crystal structure of NCX_Mj, a member of the NCX family from the archaebacterium Methanococcus jannaschii, provided insight into the atomistic details of sodium–calcium exchange. Here, we extend these findings by providing detailed functional data on purified NCX_Mj using solid supported membrane (SSM)–based electrophysiology, a powerful but unexploited tool for functional studies of electrogenic transporter proteins. We show that NCX_Mj is highly selective for Na+, whereas Ca2+ can be replaced by Mg2+ and Sr2+ and that NCX_Mj can be inhibited by divalent ions, particularly Cd2+. By directly comparing the apparent affinities of Na+ and Ca2+ for NCX_Mj with those for human NCX1, we show excellent agreement, indicating a strong functional similarity between NCX_Mj and its eukaryotic isoforms. We also provide detailed instructions to facilitate the adaption of this method to other electrogenic transporter proteins. Our findings demonstrate that NCX_Mj can serve as a model for the NCX family and highlight several possible applications for SSM-based electrophysiology.

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 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 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

Preparation and characterization of a polyethersulfone/polyaniline nanocomposite membrane for ultrafiltration and as a substrate for a gas separation membrane

RSC Advances ◽  
2015 ◽  
Vol 5 (34) ◽  
pp. 27211-27223 ◽  
Author(s):  
Shu Zhu ◽  
Mengqi Shi ◽  
Song Zhao ◽  
Zhi Wang ◽  
Jixiao Wang ◽  
...  
Keyword(s):  
Gas Separation ◽  
Woven Fabrics ◽  
Nanocomposite Membrane ◽  
Selective Layer ◽  
Gas Separation Membrane ◽  
Supported Membrane ◽  
Suitable Substrate ◽  
Separation Membrane ◽  
Polyaniline Nanocomposite

PES/PANI nanocomposite membrane displayed excellent flux and antifouling property for UF. Meanwhile, PES/PANI non-woven fabrics supported membrane performed as a suitable substrate for gas separation membrane with PVAm selective layer.

scholarly journals Functional Characterization of Bacterial NCx by Surface Supported Membrane Technology

2014 ◽  
Vol 106 (2) ◽  
pp. 581a
Author(s):  
Maria Barthmes ◽  
Jun Liao ◽  
Christian Wahl-Schott ◽  
Youxing Jiang ◽  
Andrea Brüggemann
Keyword(s):  
Functional Characterization ◽  
Membrane Technology ◽  
Supported Membrane

scholarly journals Biochemical and electrophysiological characterization of N- glycans on NMDA receptor subunits

2016 ◽  
Vol 138 (4) ◽  
pp. 546-556 ◽  
Author(s):  
Martina Kaniakova ◽  
Katarina Lichnerova ◽  
Kristyna Skrenkova ◽  
Ladislav Vyklicky ◽  
Martin Horak
Keyword(s):  
Nmda Receptor ◽  
Nmda Receptor Subunits ◽  
Electrophysiological Characterization

scholarly journals Electrophysiological Characterization of Membrane Disruption by Nanoparticles

ACS Nano ◽  
2011 ◽  
Vol 5 (5) ◽  
pp. 3599-3606 ◽  
Author(s):  
Maurits R. R. de Planque ◽  
Sara Aghdaei ◽  
Tiina Roose ◽  
Hywel Morgan
Keyword(s):  
Membrane Disruption ◽  
Electrophysiological Characterization
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