The Ionic selectivity of large protein ion channels

Els canals iònics de gran amplada i de conductància alta exerceixen una funció molt important en el cicle de vida de les cèl· lules. Permeten l'intercanvi de soluts neutres i carregats a través de la membrana cel·lular i regulen l'aportació de nutrients i la sortida de deixalles. Per a realitzar aquesta funció, els canals han de discriminar entre diferents espècies iòniques. Els canals mesoscòpics permeten el transport multiiònic passiu i generalment presenten una selectivitat moderada respecte a ions positius i negatius. Aquí revisem les metodologies més usades per a estimar quantitativament la selectivitat iònica. Entre elles, la mesura del potencial elèctric necessari per a anul·lar el corrent elèctric a través del canal en presència d'un gradient de concentració, cosa que es coneix com a potencial de corrent zero. Assenyalem els factors clau que cal tenir en compte per a una interpretació física correcta d'aquests experiments d'electrofisiologia.Large, highly conductive ion channels have a major functional role in the cell life cycle. They allow the exchange of charged and neutral solutes across the cell membrane envelope and regulate the influx of nutrients and the extrusion of waste products. To perform this function, channels must discriminate between different ionic species. Mesoscopic channels allow multiionic, passive transport and are usually moderately selective toward positive or negative ions. Here we review one of the most common approaches used for the quantitative estimation of channel selectivity: the measurement of the potential needed to get zero current across a channel in the presence of an electrolyte concentration gradient, also known as Reversal Potential. We highlight several key points that need to be addressed for a correct physical interpretation of these experiments in electrophysiology

Electrostatics Explains the Shift in VDAC Gating with Salt Activity Gradient

AbstractWe have analyzed voltage-dependent anion-selective channel (VDAC) gating on the assumption that the states occupied by the channel are determined mainly by their electrostatic energy. The voltage dependence of VDAC gating both in the presence and in the absence of a salt activity gradient was explained just by invoking electrostatic interactions. A model describing this energy in the main VDAC states has been developed. On the basis of the model, we have considered how external factors cause the redistribution of the channels among their conformational states. We propose that there is a difference in the electrostatic interaction between the voltage sensor and fixed charge within the channel when the former is located in the cis side of membrane as opposed to the trans. This could be the main cause of the shift in the probability curve. The theory describes satisfactorily the experimental data (Zizi et al., Biophys. J. 1998. 75:704–713) and explains some peculiarities of VDAC gating. The asymmetry of the probability curve was related to the apparent location of the VDAC voltage sensor in the open state. By analyzing published experimental data, we concluded that this apparent location is influenced by the diffusion potential. Also discussed is the possibility that VDAC gating at high voltage may be better described by assuming that the mobile charge consists of two parts that have to overcome different energetic barriers in the channel-closing process

SARS-CoV-2 accessory protein 7b forms homotetramers in detergent

A global pandemic is underway caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 genome, like its predecessor SARS-CoV, contains open reading frames that encode accessory proteins involved in virus-host interactions active during infection and which likely contribute to pathogenesis. One of these accessory proteins is 7b, with only 44 (SARS-CoV) and 43 (SARS-CoV-2) residues. It has one predicted transmembrane domain fully conserved, which suggests a functional role, whereas most variability is contained in the predicted cytoplasmic C-terminus. In SARS-CoV, 7b protein is expressed in infected cells, and the transmembrane domain was necessary and sufficient for Golgi localization. Also, anti-p7b antibodies have been found in the sera of SARS-CoV convalescent patients. In the present study, we have investigated the hypothesis that SARS-2 7b protein forms oligomers with ion channel activity. We show that in both SARS viruses 7b is almost completely α-helical and has a single transmembrane domain. In SDS, 7b forms various oligomers, from monomers to tetramers, but only monomers when exposed to reductants. Combination of SDS gel electrophoresis and analytical ultracentrifugation (AUC) in both equilibrium and velocity modes suggests a dimer-tetramer equilibrium, but a monomer–dimer–tetramer equilibrium in the presence of reductant. This data suggests that although disulfide-linked dimers may be present, they are not essential to form tetramers. Inclusion of pentamers or higher oligomers in the SARS-2 7b model were detrimental to fit quality. Preliminary models of this association was generated with AlphaFold2, and two alternative models were exposed to a molecular dynamics simulation in presence of a model lipid membrane. However, neither of the two models provided any evident pathway for ions. To confirm this, SARS-2 p7b was studied using Planar Bilayer Electrophysiology. Addition of p7b to model membranes produced occasional membrane permeabilization, but this was not consistent with bona fide ion channels made of a tetrameric assembly of α-helices

Corneal relaxation time estimation as a function of tear oxygen tension in human cornea during contact lens wear

[EN] The purpose is to estimate the oxygen diffusion coefficient and the relaxation time of the cornea with respect to the oxygen tension at the cornea-tears interface. Both findings are discussed. From the experimental data provided by Bonanno et al., the oxygen tension measurements in vivo for human cornea-tears-contact lens (CL), the relaxation time of the cornea, and their oxygen diffusion coefficient were obtained by numerical calculation using the Monod-kinetic model. Our results, considering the relaxation time of the cornea, observe a different behavior. At the time less than 8 s, the oxygen diffusivity process is upper-diffusive, and for the relaxation time greater than 8 s, the oxygen diffusivity process is lower-diffusive. Both cases depend on the partial pressure of oxygen at the entrance of the cornea. The oxygen tension distribution in the cornea-tears interface is separated into two different zones: one for conventional hydrogels, which is located between 6 and 75 mmHg, with a relaxation time included between 8 and 19 s, and the other zone for silicone hydrogel CLs, which is located at high oxygen tension, between 95 and 140 mmHg, with a relaxation time in the interval of 1.5-8 s. It is found that in each zone, the diffusion coefficient varies linearly with the oxygen concentration, presenting a discontinuity in the transition of 8 s. This could be interpreted as an aerobic-to-anaerobic transition. We attribute this behavior to the coupling formalism between oxygen diffusion and biochemical reactions to produce adenosine triphosphate.Contract grant sponsor: Dirección General de Asuntos del Personal Académico, Universidad Nacional Autónoma de México; contract grant number: UNAM-DGAPA-PAPIIT projects IG 100618 and IN-114818 Contract grant sponsor: Secretaría de Estado de Investigación, Desarrollo e Innovación; contract grant number: ENE/2015-69203-RDel Castillo, LF.; Ramírez-Calderón, JG.; Del Castillo, RM.; Aguilella-Arzo, M.; Compañ Moreno, V. (2020). Corneal relaxation time estimation as a function of tear oxygen tension in human cornea during contact lens wear. Journal of Biomedical Materials Research Part B Applied Biomaterials. 108(1):14-21. https://doi.org/10.1002/jbm.b.34360S14211081Freeman, R. D. (1972). Oxygen consumption by the component layers of the cornea. The Journal of Physiology, 225(1), 15-32. doi:10.1113/jphysiol.1972.sp009927CHALMERS, R. L., McNALLY, J. 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A., & Mandell, R. B. (1970). Critical Oxygen Tension at the Corneal Surface. Archives of Ophthalmology, 84(4), 505-508. doi:10.1001/archopht.1970.00990040507021Giasson, C., & Bonanno, J. A. (1995). Acidification of rabbit corneal endothelium during contact lens wearin vitro. Current Eye Research, 14(4), 311-318. doi:10.3109/02713689509033531Riley, M. V. (1969). Glucose and oxygen utilization by the rabbit cornea. Experimental Eye Research, 8(2), 193-200. doi:10.1016/s0014-4835(69)80031-xFrahm, B., Lane, P., M�rkl, H., & P�rtner, R. (2003). Improvement of a mammalian cell culture process by adaptive, model-based dialysis fed-batch cultivation and suppression of apoptosis. Bioprocess and Biosystems Engineering, 26(1), 1-10. doi:10.1007/s00449-003-0335-zCompañ, V., Aguilella-Arzo, M., Del Castillo, L. F., Hernández, S. I., & Gonzalez-Meijome, J. M. (2016). Analysis of the application of the generalized monod kinetics model to describe the human corneal oxygen-consumption rate during soft contact lens wear. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 105(8), 2269-2281. doi:10.1002/jbm.b.33764Bonanno, J. A., Clark, C., Pruitt, J., & Alvord, L. (2009). Tear Oxygen Under Hydrogel and Silicone Hydrogel Contact Lenses in Humans. Optometry and Vision Science, 86(8), E936-E942. doi:10.1097/opx.0b013e3181b2f582Chhabra, M., Prausnitz, J. M., & Radke, C. J. (2008). Diffusion and Monod kinetics to determine in vivo human corneal oxygen-consumption rate during soft contact-lens wear. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 90B(1), 202-209. doi:10.1002/jbm.b.31274Chhabra, M., Prausnitz, J. M., & Radke, C. J. (2009). Modeling Corneal Metabolism and Oxygen Transport During Contact Lens Wear. Optometry and Vision Science, 86(5), 454-466. doi:10.1097/opx.0b013e31819f9e70Larrea, X., & Bu¨chler, P. (2009). A Transient Diffusion Model of the Cornea for the Assessment of Oxygen Diffusivity and Consumption. Investigative Opthalmology & Visual Science, 50(3), 1076. doi:10.1167/iovs.08-2479Alvord, L. A., Hall, W. J., Keyes, L. D., Morgan, C. F., & Winterton, L. C. (2007). Corneal Oxygen Distribution With Contact Lens Wear. Cornea, 26(6), 654-664. doi:10.1097/ico.0b013e31804f5a22Del Castillo, L. F., da Silva, A. R. F., Hernández, S. I., Aguilella, M., Andrio, A., Mollá, S., & Compañ, V. (2015). Diffusion and Monod kinetics model to determine in vivo human corneal oxygen-consumption rate during soft contact lens wear. Journal of Optometry, 8(1), 12-18. doi:10.1016/j.optom.2014.06.002Chandel, N. S., Budinger, G. R. S., Choe, S. H., & Schumacker, P. T. (1997). Cellular Respiration during Hypoxia. Journal of Biological Chemistry, 272(30), 18808-18816. doi:10.1074/jbc.272.30.18808Leung, B. K., Bonanno, J. A., & Radke, C. J. (2011). Oxygen-deficient metabolism and corneal edema. Progress in Retinal and Eye Research, 30(6), 471-492. doi:10.1016/j.preteyeres.2011.07.001Chhabra, M., Prausnitz, J. M., & Radke, C. J. (2008). Polarographic Method for Measuring Oxygen Diffusivity and Solubility in Water-Saturated Polymer Films:  Application to Hypertransmissible Soft Contact Lenses. Industrial & Engineering Chemistry Research, 47(10), 3540-3550. doi:10.1021/ie071071aCompañ, V., Andrio, A., López-Alemany, A., Riande, E., & Refojo, M. F. (2002). Oxygen permeability of hydrogel contact lenses with organosilicon moieties. Biomaterials, 23(13), 2767-2772. doi:10.1016/s0142-9612(02)00012-1Gonzalez-Meijome, J. M., Compañ-Moreno, V., & Riande, E. (2008). Determination of Oxygen Permeability in Soft Contact Lenses Using a Polarographic Method:  Estimation of Relevant Physiological Parameters. Industrial & Engineering Chemistry Research, 47(10), 3619-3629. doi:10.1021/ie071403bCompa�, V., L�pez, M. L., Andrio, A., L�pez-Alemany, A., & Refojo, M. F. (1999). 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(2003). Thickness of the Pre- and Post–Contact Lens Tear Film Measured In Vivo by Interferometry. Investigative Opthalmology & Visual Science, 44(1), 68. doi:10.1167/iovs.02-0377Compañ, V., Aguilella-Arzo, M., Edrington, T. B., & Weissman, B. A. (2016). Modeling Corneal Oxygen with Scleral Gas Permeable Lens Wear. Optometry and Vision Science, 93(11), 1339-1348. doi:10.1097/opx.0000000000000988Compañ, V., Aguilella-Arzo, M., & Weissman, B. A. (2017). Corneal Equilibrium Flux as a Function of Corneal Surface Oxygen Tension. Optometry and Vision Science, 94(6), 672-679. doi:10.1097/opx.0000000000001083Papas, E. B., & Sweeney, D. F. (2016). Interpreting the corneal response to oxygen: Is there a basis for re-evaluating data from gas-goggle studies? Experimental Eye Research, 151, 222-226. doi:10.1016/j.exer.2016.08.019Alentiev, A. Y., Shantarovich, V. P., Merkel, T. C., Bondar, V. I., Freeman, B. D., & Yampolskii, Y. P. (2002). 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Amphiphilic COSAN and I2-COSAN crossing synthetic lipid membranes: planar bilayers and liposomes

The boron-rich cobaltabisdicarbollide (COSAN) and its 8,80-I2 derivative (I2-COSAN), both of purely inorganic nature, are shown to cross through synthetic lipid membranes. These results reveal unexpected properties at the interface of biological and synthetic membranes

Comments to paper entitled "Predicting scleral GP lens entrapped tear layer oxygen tensions"

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Severe acute respiratory syndrome coronavirus E protein transports calcium ions and activates the NLRP3 inflammasome

Severe acute respiratory syndrome coronavirus (SARS-CoV) envelope (E) protein is a viroporin involved in virulence. E protein ion channel (IC) activity is specifically correlated with enhanced pulmonary damage, edema accumulation and death. IL-1β driven proinflammation is associated with those pathological signatures, however its link to IC activity remains unknown. In this report, we demonstrate that SARS-CoV E protein forms protein–lipid channels in ERGIC/Golgi membranes that are permeable to calcium ions, a highly relevant feature never reported before. Calcium ions together with pH modulated E protein pore charge and selectivity. Interestingly, E protein IC activity boosted the activation of the NLRP3 inflammasome, leading to IL-1β overproduction. Calcium transport through the E protein IC was the main trigger of this process. These findings strikingly link SARS-CoV E protein IC induced ionic disturbances at the cell level to immunopathological consequences and disease worsening in the infected organism