A small viral potassium ion channel with an inherent inward rectification

Eckert, Denise; Schulze, Tobias; Stahl, Julian; Rauh, Oliver; Etten, James L. Van; Hertel, Brigitte; Schroeder, Indra; Moroni, Anna; Thiel, Gerhard

doi:10.1080/19336950.2019.1605813

Cited by 6 publications

(2 citation statements)

References 47 publications

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“…The relative polarization of the mitochondrial membrane was measured as described previously [ 26 ] using the accumulation of the fluorescent mitochondrial marker tetramethylrhodamine methyl ester perchlorate (TMRM) as an indirect measure of the mitochondrial membrane potential. HEK293 cells were measured >18 h after transfection with the Kesv channel plus GFP tag.…”

Section: Methodsmentioning

confidence: 99%

Light-Regulated Transcription of a Mitochondrial-Targeted K+ Channel

Engel

Winterstein

Kithil

et al. 2020

Cells

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The inner membranes of mitochondria contain several types of K+ channels, which modulate the membrane potential of the organelle and contribute in this way to cytoprotection and the regulation of cell death. To better study the causal relationship between K+ channel activity and physiological changes, we developed an optogenetic platform for a light-triggered modulation of K+ conductance in mitochondria. By using the light-sensitive interaction between cryptochrome 2 and the regulatory protein CIB1, we can trigger the transcription of a small and highly selective K+ channel, which is in mammalian cells targeted into the inner membrane of mitochondria. After exposing cells to very low intensities (≤0.16 mW/mm2) of blue light, the channel protein is detectable as an accumulation of its green fluorescent protein (GFP) tag in the mitochondria less than 1 h after stimulation. This system allows for an in vivo monitoring of crucial physiological parameters of mitochondria, showing that the presence of an active K+ channel causes a substantial depolarization compatible with the effect of an uncoupler. Elevated K+ conductance also results in a decrease in the Ca2+ concentration in the mitochondria but has no impact on apoptosis.

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Section: Methodsmentioning

confidence: 99%

Light-Regulated Transcription of a Mitochondrial-Targeted K+ Channel

Engel

Winterstein

Kithil

et al. 2020

Cells

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“…In essence, the semipermeable lipid-based plasma membrane acts as an electrical insulator, but also as a capacitor that can accumulate charge, while specialized passages (ion channels, pumps, connexins/gap junctions, and solute carriers) regulate ion flow from one side to the other, altering the voltage of the cell ( Figure 1 A). All cell types form ionic gradients across their cell membranes because channels exist throughout all living organisms in all domains of life, including plants, fungi, and bacteria [ 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 ]. Thus, ion regulation and the resulting bioelectricity are considered essential properties of living cells across evolution, and their innate properties can be used for cellular communication [ 25 , 26 ].…”

Section: Introductionmentioning

confidence: 99%

Bioelectricity in Developmental Patterning and Size Control: Evidence and Genetically Encoded Tools in the Zebrafish Model

Silic

Zhang

2023

Cells

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Developmental patterning is essential for regulating cellular events such as axial patterning, segmentation, tissue formation, and organ size determination during embryogenesis. Understanding the patterning mechanisms remains a central challenge and fundamental interest in developmental biology. Ion-channel-regulated bioelectric signals have emerged as a player of the patterning mechanism, which may interact with morphogens. Evidence from multiple model organisms reveals the roles of bioelectricity in embryonic development, regeneration, and cancers. The Zebrafish model is the second most used vertebrate model, next to the mouse model. The zebrafish model has great potential for elucidating the functions of bioelectricity due to many advantages such as external development, transparent early embryogenesis, and tractable genetics. Here, we review genetic evidence from zebrafish mutants with fin-size and pigment changes related to ion channels and bioelectricity. In addition, we review the cell membrane voltage reporting and chemogenetic tools that have already been used or have great potential to be implemented in zebrafish models. Finally, new perspectives and opportunities for bioelectricity research with zebrafish are discussed.

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Cell-Free Protein Synthesis: A Promising Option for Future Drug Development

et al. 2020

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Proteins are the main source of drug targets and some of them possess therapeutic potential themselves. Among them, membrane proteins constitute approximately 50% of the major drug targets. In the drug discovery pipeline, rapid methods for producing different classes of proteins in a simple manner with high quality are important for structural and functional analysis. Cell-free systems are emerging as an attractive alternative for the production of proteins due to their flexible nature without any cell membrane constraints. In a bioproduction context, open systems based on cell lysates derived from different sources, and with batch-to-batch consistency, have acted as a catalyst for cell-free synthesis of target proteins. Most importantly, proteins can be processed for downstream applications like purification and functional analysis without the necessity of transfection, selection, and expansion of clones. In the last 5 years, there has been an increased availability of new cell-free lysates derived from multiple organisms, and their use for the synthesis of a diverse range of proteins. Despite this progress, major challenges still exist in terms of scalability, cost effectiveness, protein folding, and functionality. In this review, we present an overview of different cell-free systems derived from diverse sources and their application in the production of a wide spectrum of proteins. Further, this article discusses some recent progress in cell-free systems derived from Chinese hamster ovary and Sf21 lysates containing endogenous translocationally active microsomes for the synthesis of membrane proteins. We particularly highlight the usage of internal ribosomal entry site sequences for more efficient protein production, and also the significance of site-specific incorporation of non-canonical amino acids for labeling applications and creation of antibody drug conjugates using cell-free systems. We also discuss strategies to overcome the major challenges involved in commercializing cell-free platforms from a laboratory level for future drug development. Key PointsCell-free protein synthesis has the potential to become an alternative method for the rapid and highly parallel expression of a diverse range of proteins.Cell-free systems utilize the translation machinery of the cells, bypassing the constraints of the cell membrane and thus offer openness and configurational flexibility even for the synthesis of difficult-to-express proteins.In comparison to traditional approaches, cell-free systems can be time-saving, from initial synthesis to downstream applications.

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A small viral potassium ion channel with an inherent inward rectification

Cited by 6 publications

References 47 publications

Light-Regulated Transcription of a Mitochondrial-Targeted K+ Channel

Light-Regulated Transcription of a Mitochondrial-Targeted K+ Channel

Bioelectricity in Developmental Patterning and Size Control: Evidence and Genetically Encoded Tools in the Zebrafish Model

Cell-Free Protein Synthesis: A Promising Option for Future Drug Development

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