title: Computational simulations of asymmetric fluxes of large molecules through gap junction channel pores.
abstract: Gap junction channels are formed out of connexin isoforms, which enable molecule and ion selective diffusion amongst neighboring cells. HeLa cells expressing distinct connexins (Cx) allow the formation of heterotypic channels, where we observed a molecular charge-independent preferential flux of large fluorescent molecules in the Cx45 to Cx43 direction. We hypothesize that the pore's shape is a significant factor along-side charge and transjunctional voltages for this asymmetric flux. To test this hypothesis, we developed a 3D computational model simulating Brownian diffusion of large molecules in a gap junction channel pore. The basic pore contour was derived from x-ray crystallographic structures of Cx43 and Cx26 and approximated using basic geometric shapes. Lucifer yellow dye molecules and cesium counter-ions were modeled as spheres using their respective Stokes radii. Our simulation results from simple diffusion and constant concentration gradient experiments showed that only charged particles yield asymmetric fluxes in heterotypic pores. While increasing the inner mouth size resulted in a near-quadratic rise in flux, the rise was asymptotic for outer mouth radii increase. Probability maps and average force per particle per pore section explain the asymmetric flux with variation in pore shape. Furthermore, the simulation results are in agreement with our in vitro experimental results with HeLa cells in Cx43-Cx45 heterotypic configurations. The presence of asymmetric fluxes can help us to understand effects of the molecular structure of the pore and predict potential differences in vivo.
authors: Department of Bioengineering, University of Utah, USA; Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah, USA. Electronic address: abhijit.mondal@utah.edu.
authors: Abhijit
authors: A
authors: Mondal
authors: Department of Biology, University of Utah, USA.
authors: Daniel A
authors: DA
authors: Appadurai
authors: Department of Cardiology, University of Washington, USA.
authors: Nazem W
authors: NW
authors: Akoum
authors: Department of Bioengineering, University of Utah, USA; Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah, USA.
authors: Frank B
authors: FB
authors: Sachse
authors: Department of Bioengineering, University of Utah, USA; Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah, USA. Electronic address: alonso.moreno@utah.edu.
authors: Alonso P
authors: AP
authors: Moreno
mesh_terms: D000072259:Connexin 26; D018031:Connexin 43 / Q000737:chemistry* / Q000378:metabolism; D017630:Connexins / Q000737:chemistry* / Q000378:metabolism; D017629:Gap Junctions / Q000737:chemistry* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular
chemical_list: D018031:Connexin 43; D017630:Connexins; C000607042:GJB2 protein, human; D000072259:Connexin 26
keywords: Asymmetric flux; Brownian dynamics simulation; Computational model; Connexins; Gap junctions; Heterotypic