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Full-Text Articles in Life Sciences
The Hydrophobic Residues In Amino Terminal Domains Of Cx46 And Cx50 Are Important For Their Gap Junction Channel Ion Permeation And Gating, Roa'a Jaradat
Electronic Thesis and Dissertation Repository
Connexins are ubiquitous transmembrane proteins that assemble to form intercellular channels, called gap junctions (GJs). The amino terminal (NT) domain and residues within this domain have been shown to be important for both Vj-gating and single channel conductance (γj) of several GJs including Cx46 and Cx50. High-resolution structures of Cx46 and Cx50 GJs were recently resolved showing that NT domain folds into the inner pore where hydrophobic residues are packed against M1/2 domains stabilizing open-state conformation. We studied functional properties of GJs formed by several point variants at NT (Cx46 L10I, N13E, A14V, Q15N and Cx50 …
The Amino Terminal Domains Of Sheep Cx46 Or Cx50 Determine Their Gap Junction Channel Open Stability And Unitary Channel Conductance, Benny Yue
Electronic Thesis and Dissertation Repository
Connexins form intercellular communication channels known as gap junctions (GJs), which are found throughout the vertebrate species. GJs formed by different connexins harbor unique channel properties that have not been fully defined. High-resolution structures of native Cx46 and Cx50 GJs from sheep (sCx46 and sCx50) were recently resolved. Molecular dynamics studies identified the NT domains, especially the 9th position, as key determinants in the differences of energetic barrier to K+ permeation in sCx46 and sCx50 GJs. We studied functional properties of GJs formed by sCx46, sCx50, NT domain swapped chimeras (sCx46-50NT and sCx50-46NT), and point variants at the …
Heterotypic Docking Compatibility Of Human Connexin 37 With Other Vascular Connexins, Nicholas Kim
Heterotypic Docking Compatibility Of Human Connexin 37 With Other Vascular Connexins, Nicholas Kim
Electronic Thesis and Dissertation Repository
Gap junction (GJ) channels provide direct intercellular communication. A GJ channel consists of two docked hemichannels and each hemichannel is a hexamer of six connexins. Human vascular connexins (Cx37, Cx40, Cx43, and Cx45) can form various types of gap junction channels to synchronize vasodilation/constriction to control local circulation. Most of our knowledge on heterotypic GJs of these vascular connexins comes from studies on rodent connexins. However it is not clear if these human connexins can also form heterotypic GJs. The present study used an in vitro expression system to investigate the coupling status and GJ properties of human heterotypic Cx37/Cx40, …
Functional Characteristics Of Four Novel Lone Atrial Fibrillation-Linked Connexin40 Mutants, Mahmoud Noureldin
Functional Characteristics Of Four Novel Lone Atrial Fibrillation-Linked Connexin40 Mutants, Mahmoud Noureldin
Electronic Thesis and Dissertation Repository
Atrial fibrillation (AF) is the most common form of cardiac arrhythmia. Recently, four novel heterozygous Cx40 mutations, K107R, L223M, Q236H, and I257L were identified in 4 of 310 unrelated AF patients. To study possible alterations associated with these mutants, we studied their localization and function using gap junction (GJ)-deficient model cells. Cell pairs expressing Q236H alone or together with wildtype Cx43 showed a significantly lower coupling conductance. Impaired GJ function and dominant negative action on Cx43 of this mutant are consistent with previous findings on the majority of AF-linked Cx40 mutants. The remaining three novel AF-linked mutants did not show …
Putative Pore-Lining Residues And Intracellular Magnesium Influence Connexin50 Unitary Gap Junction Channel Conductance, Swathy Sudhakar
Putative Pore-Lining Residues And Intracellular Magnesium Influence Connexin50 Unitary Gap Junction Channel Conductance, Swathy Sudhakar
Electronic Thesis and Dissertation Repository
Gap junction channels (GJs) are oligomers of connexins forming channels linking neighbouring cells. GJs formed by different connexins show distinct unitary channel conductance (γj), transjunctional-voltage dependent gating (Vj-gating) properties, and magnesium modulation. We investigate here how pore-lining residues may influence these GJ properties. We mutated putative pore-lining residues in Cx50 GJ (G8E/G46E/V53E individually or in combination) to investigate γj, Vj-gating, and magnesium modulation. GJs formed by combination mutants (G8E-G46E, G46E-V53E, and G8E-G46E-V53E) were functional and showed a significantly increased γj with little change in Vj-gating, while G8E-V53E mutants failed …
Engineered Cx26 Variant Established Functional Heterotypic Cx26/Cx43 And Cx26/Cx40 Gap Junction Channels, Levent Berk Karademir
Engineered Cx26 Variant Established Functional Heterotypic Cx26/Cx43 And Cx26/Cx40 Gap Junction Channels, Levent Berk Karademir
Electronic Thesis and Dissertation Repository
Two hexameric connexin hemichannels dock together to from a whole gap junction (GJ) channel. The mechanisms of docking specificity in forming homotypic and heterotypic GJ channels are not fully clear. To reveal the key differences between Cx26 and Cx43 (or Cx40) in their docking residues, we aligned and analyzed ten well studied connexin sequences. Five of them are docking compatible with Cx26 and the rest (including Cx43 and Cx40) are not. According to Cx26 crystal structure at the docking interface, we identified two putative docking residues on the second extracellular domain (E2) that are well conserved within docking compatible connexins, …
Mutations At The 46th Residue Change Properties Of Cx50 Gap Junction Channels, Xiaoling Tong
Mutations At The 46th Residue Change Properties Of Cx50 Gap Junction Channels, Xiaoling Tong
Electronic Thesis and Dissertation Repository
The interface of the first transmembrane domain and the first extracellular loop (TM1/E1 border) in several gap junction (GJ) channels is known to line a portion of the pore and plays an important role in determining GJ channel properties. By introduction of a charged residue into this domain of Cx50, the resultant mutant channels showed drastically altered unitary conductance (γj) and transjunctional voltage-dependent gating (Vj-gating). Specifically G46D and G46E increased the Cx50 γj from 201 to 256 and 293 pS, respectively and G46K channel showed a decreased γj of only 20 pS. Moreover, in …
Presence Of A Cs-Resistant Transient Outward Current Which Distorts Ica Measurements In Guinea-Pig Single Ventricular Cells, Georges Christé, Brigitte Delachapelle, Fouad Lemtiri-Chlieh, Carlos Ojeda
Presence Of A Cs-Resistant Transient Outward Current Which Distorts Ica Measurements In Guinea-Pig Single Ventricular Cells, Georges Christé, Brigitte Delachapelle, Fouad Lemtiri-Chlieh, Carlos Ojeda
fouad Lemtiri-Chlieh
To correctlyr analyse the physical properties of ion channels and the actions of drugs upon them, it is important that the membrane conducts only the ionic species believed permeant. In guinea-pig ventricular cells it has been assumed that only ICaL flows in the potential range -50 to +60 mV in the presence of intra- and extracellular Cs (> 20 mM). We report here that this is not the case due to the presence of a Cs-insensitive, transient outward current.
Measurements Of Cytosolic Free Ca In Cultured Rat Neonate And Adult Guinea-Pig Ventricular Cardiac Myocytes, Jean-Claude Bernengo, Fouad Lemtiri-Chlieh, Carlos Ojeda, Nadine Pltonoff
Measurements Of Cytosolic Free Ca In Cultured Rat Neonate And Adult Guinea-Pig Ventricular Cardiac Myocytes, Jean-Claude Bernengo, Fouad Lemtiri-Chlieh, Carlos Ojeda, Nadine Pltonoff
fouad Lemtiri-Chlieh
Cytosolic free Ca was measured with Indo-1 (Grynkiewicz et al. 1985) as free acid (in guinea-pig cells) or as the ester (in rat neonate cells). Using image analysis we obtained concentration maps at 40 ms intervals at two wavelengths (Cannell et al. 1986). In rat neonate cells, at rest and during contraction, calcium was apparently non-uniformly distributed, in contrast to guinea-pig adult cells. Using a dual PM spectroscopic system adapted to an inverted microscope we found: (1) the conversion of Indo-AM to Indo- 1 in the cytosol is not complete (< 40% contribution to the total light); (2) rat neonate cells buffer Ca (at rest 130-170 nM) only for [Ca]. between 1 and 3 mM, whereas in adult guinea-pig cells the range extended to 10 mM; (3) during contraction [Ca]i rose from 200 nm to no more than 1 microM. Digitoxin increases basal and peak [Ca]i; (4) during spontaneous contractions, in guinea-pig cells, [Ca]i rises from 70 to 200 nm when [Ca]o = 0, and from 170 nm to about 1 microM when [Ca]o = 2 mM; (5) in voltage-clamp conditions the rise in [Ca]i does not exceed 2 microM and oscillations occur in the presence of forskolin. When loading with Indo-1, cells frequently cease to contract (Powell et al. 1988) and the variations in [Ca]i are then very small (about 200 nm at peak ICa). In both cases these values are small compared to the total charge crossing the membrane.