Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Microarray-based screening system identifies temperature-controlled activity of Connexin 26 that is distorted by mutations.
Wang H
,
Stahl F
,
Scheper T
,
Steffens M
,
Warnecke A
,
Zeilinger C
.
???displayArticle.abstract???
Here, we show that human Connexin 26 (hCx26 or Cx26WT) hemichannel opening rapidly enables the transport of small molecules when triggered by temperature and by compensation of the Ca2+ blockade with EDTA. Point mutations within Cx26 were analysed by a novel optical microarray-based Lucifer Yellow uptake assay or by two electrode voltage clamp (TEVC) on frog oocytes to monitor simultaneous activities of channel proteins. Point mutations L90P, F161S, R184P or K188N influenced the temperature-dependent activity drastically. Since several mutations blocked trafficking, the temperature-dependent activity of the recombinant synthesized and purified wild-type Cx26WT and Cx26K188N hemichannel was tested by liposome flux assay (LFA) and on a microarray-based Lucifer Yellow uptake assay under warm conditions (>30 °C). The data from TEVC measurements and dye flux experiments showed that the mutations gave no or only a weak activity at increased temperature (>30 °C). We conclude that the position K188 in the Cx26WT forms a temperature-sensitive salt bridge with E47 whereas the exchange to K188N destabilizes the network loop- gating filter, which was recently identified as a part of the flexible Ca2+ binding site. We assume that the temperature sensitivity of Cx26 is required to protect cells from uncontrolled release or uptake activities through Cx26 hemichannels.
Aasen,
Connexins: Synthesis, Post-Translational Modifications, and Trafficking in Health and Disease.
2018, Pubmed
Aasen,
Connexins: Synthesis, Post-Translational Modifications, and Trafficking in Health and Disease.
2018,
Pubmed
Ambrosi,
Analysis of trafficking, stability and function of human connexin 26 gap junction channels with deafness-causing mutations in the fourth transmembrane helix.
2013,
Pubmed
,
Xenbase
Bargiello,
Gating of Connexin Channels by transjunctional-voltage: Conformations and models of open and closed states.
2018,
Pubmed
Bargiello,
Voltage-dependent conformational changes in connexin channels.
2012,
Pubmed
,
Xenbase
Bennett,
An electrostatic mechanism for Ca(2+)-mediated regulation of gap junction channels.
2016,
Pubmed
Bukauskas,
Gap junction channel gating.
2004,
Pubmed
Burnstock,
Purinergic signaling in healthy and diseased skin.
2012,
Pubmed
Chan,
GJB2-associated hearing loss: systematic review of worldwide prevalence, genotype, and auditory phenotype.
2014,
Pubmed
Chang,
Timed conditional null of connexin26 in mice reveals temporary requirements of connexin26 in key cochlear developmental events before the onset of hearing.
2015,
Pubmed
Chen,
Deafness induced by Connexin 26 (GJB2) deficiency is not determined by endocochlear potential (EP) reduction but is associated with cochlear developmental disorders.
2014,
Pubmed
Conti,
Transgenic mice with a reduced core body temperature have an increased life span.
2006,
Pubmed
Danzl,
Early inhibition of endothelial retinoid uptake upon myocardial infarction restores cardiac function and prevents cell, tissue, and animal death.
2019,
Pubmed
Delvaeye,
Therapeutic Targeting of Connexin Channels: New Views and Challenges.
2018,
Pubmed
Edelheit,
Simple and efficient site-directed mutagenesis using two single-primer reactions in parallel to generate mutants for protein structure-function studies.
2009,
Pubmed
Evans,
Gap junctions: structure and function (Review).
2002,
Pubmed
Fetoni,
Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway.
2018,
Pubmed
García Gómez-Heras,
Purkinje fibers after myocardial ischemia-reperfusion.
2015,
Pubmed
Gerido,
Aberrant hemichannel properties of Cx26 mutations causing skin disease and deafness.
2007,
Pubmed
,
Xenbase
Ito,
Thermodynamic implications of high Q 10 of thermo-TRP channels in living cells.
2015,
Pubmed
Jonczyk,
Development of living cell microarrays using non-contact micropipette printing.
2016,
Pubmed
Kecskeméti,
Analysis of GJB2 mutations and the clinical manifestation in a large Hungarian cohort.
2018,
Pubmed
Klein,
Thermosensitive transient receptor potential (TRP) channel agonists and their role in mechanical, thermal and nociceptive sensations as assessed using animal models.
2015,
Pubmed
Kniggendorf,
Temperature-sensitive gating of hCx26: high-resolution Raman spectroscopy sheds light on conformational changes.
2014,
Pubmed
Kwon,
Molecular dynamics simulations of the Cx26 hemichannel: evaluation of structural models with Brownian dynamics.
2011,
Pubmed
,
Xenbase
Kwon,
Molecular dynamics simulations of the Cx26 hemichannel: insights into voltage-dependent loop-gating.
2012,
Pubmed
Laird,
Therapeutic strategies targeting connexins.
2018,
Pubmed
Laird,
Life cycle of connexins in health and disease.
2006,
Pubmed
Lang,
Calcium-Dependent Arrhythmogenic Foci Created by Weakly Coupled Myocytes in the Failing Heart.
2017,
Pubmed
Lefebvre,
Connexins, hearing and deafness: clinical aspects of mutations in the connexin 26 gene.
2000,
Pubmed
Liang,
Cell degeneration is not a primary causer for Connexin26 (GJB2) deficiency associated hearing loss.
2012,
Pubmed
Lopez,
Insights on the mechanisms of Ca(2+) regulation of connexin26 hemichannels revealed by human pathogenic mutations (D50N/Y).
2013,
Pubmed
,
Xenbase
Mammano,
Inner Ear Connexin Channels: Roles in Development and Maintenance of Cochlear Function.
2019,
Pubmed
Mani,
Functional consequences of novel connexin 26 mutations associated with hereditary hearing loss.
2009,
Pubmed
Mei,
A deafness mechanism of digenic Cx26 (GJB2) and Cx30 (GJB6) mutations: Reduction of endocochlear potential by impairment of heterogeneous gap junctional function in the cochlear lateral wall.
2017,
Pubmed
Moscato,
Connexin 26 Expression in Mammalian Cardiomyocytes.
2018,
Pubmed
Myers,
Structure of native lens connexin 46/50 intercellular channels by cryo-EM.
2018,
Pubmed
Naz,
Genetic causes of moderate to severe hearing loss point to modifiers.
2017,
Pubmed
Neijssen,
Cross-presentation by intercellular peptide transfer through gap junctions.
2005,
Pubmed
Ngezahayo,
Inactivation of expressed and conducting rCx46 hemichannels by phosphorylation.
1998,
Pubmed
,
Xenbase
Oh,
Voltage regulation of connexin channel conductance.
2015,
Pubmed
Petit,
Molecular genetics of hearing loss.
2001,
Pubmed
Qu,
Accessibility of cx46 hemichannels for uncharged molecules and its modulation by voltage.
2004,
Pubmed
,
Xenbase
Ripps,
Properties of connexin26 hemichannels expressed in Xenopus oocytes.
2004,
Pubmed
,
Xenbase
Santos-Miranda,
Effects of temperature on transjunctional voltage-dependent gating kinetics in Cx45 and Cx40 gap junction channels.
2019,
Pubmed
Srinivas,
Human diseases associated with connexin mutations.
2018,
Pubmed
Steffens,
Regulation of connexons composed of human connexin26 (hCx26) by temperature.
2008,
Pubmed
,
Xenbase
Su,
Novel cell-free high-throughput screening method for pharmacological tools targeting K+ channels.
2016,
Pubmed
Sánchez,
Differentially altered Ca2+ regulation and Ca2+ permeability in Cx26 hemichannels formed by the A40V and G45E mutations that cause keratitis ichthyosis deafness syndrome.
2010,
Pubmed
,
Xenbase
Taki,
Roles of aberrant hemichannel activities due to mutant connexin26 in the pathogenesis of KID syndrome.
2018,
Pubmed
Tamames,
Theoretical Evaluation and Experimental Validation of Localized Therapeutic Hypothermia Application to Preserve Residual Hearing After Cochlear Implantation.
2018,
Pubmed
Thönnissen,
Human connexin26 (GJB2) deafness mutations affect the function of gap junction channels at different levels of protein expression.
2002,
Pubmed
Wang,
Adenosine triphosphate acts as a paracrine signaling molecule to reduce the motility of T cells.
2014,
Pubmed
Wang,
Children with GJB2 gene mutations have various audiological phenotypes.
2018,
Pubmed
Wingard,
Cellular and Deafness Mechanisms Underlying Connexin Mutation-Induced Hearing Loss - A Common Hereditary Deafness.
2015,
Pubmed
Xiao,
Thermosensation and longevity.
2015,
Pubmed
Zeilinger,
Length of C-terminus of rCx46 influences oligomerization and hemichannel properties.
2005,
Pubmed
,
Xenbase
Zhang,
Conserved glycine at position 45 of major cochlear connexins constitutes a vital component of the Ca²⁺ sensor for gating of gap junction hemichannels.
2013,
Pubmed
Zhu,
Connexin26 (GJB2) deficiency reduces active cochlear amplification leading to late-onset hearing loss.
2015,
Pubmed
Zong,
Progressive age-dependence and frequency difference in the effect of gap junctions on active cochlear amplification and hearing.
2017,
Pubmed
Zonta,
Cues to Opening Mechanisms From in Silico Electric Field Excitation of Cx26 Hemichannel and in Vitro Mutagenesis Studies in HeLa Transfectans.
2018,
Pubmed
Zonta,
Role of gamma carboxylated Glu47 in connexin 26 hemichannel regulation by extracellular Ca²⁺: insight from a local quantum chemistry study.
2014,
Pubmed