The sialic acid component of the beta1 subunit modulates voltage-gated sodium channel function.
Voltage-gated sodium channels (Nav) are responsible for initiation and propagation of nerve, skeletal muscle, and cardiac action potentials. Nav are composed of a pore-forming alpha subunit and often one to several modulating beta subunits. Previous work showed that terminal sialic acid residues attached to alpha subunits affect channel gating. Here we show that the fully sialylated beta1 subunit induces a uniform, hyperpolarizing shift in steady state and kinetic gating of the cardiac and two neuronal alpha subunit isoforms. Under conditions of reduced sialylation, the beta1-induced gating effect was eliminated. Consistent with this, mutation of beta1 N-glycosylation sites abolished all effects of beta1 on channel gating. Data also suggest an interaction between the cis effect of alpha sialic acids and the trans effect of beta1 sialic acids on channel gating. Thus, beta1 sialic acids had no effect gating on the of the heavily glycosylated skeletal muscle alpha subunit. However, when glycosylation of the skeletal muscle alpha subunit was reduced through chimeragenesis such that alpha sialic acids did not impact gating, beta1 sialic acids caused a significant hyperpolarizing shift in channel gating. Together, the data indicate that beta1 N-linked sialic acids can modulate Nav gating through an apparent saturating electrostatic mechanism. A model is proposed in which a spectrum of differentially sialylated Nav can directly modulate channel gating, thereby impacting cardiac, skeletal muscle, and neuronal excitability.
Johnson
Daniel
D
Department of Physiology & Biophysics and Program in Neuroscience, University of South Florida College of Medicine, Tampa, Florida 33612, USA.
Montpetit
Marty L
ML
Stocker
Patrick J
PJ
Bennett
Eric S
ES
D000818:Animals; D016466:CHO Cells; D002241:Carbohydrates / Q000737:chemistry; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D004594:Electrophysiology; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D008564:Membrane Potentials; D008954:Models, Biological; D018482:Muscle, Skeletal / Q000378:metabolism; D016296:Mutagenesis; D009154:Mutation; D009206:Myocardium / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000737:chemistry* / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D020033:Protein Isoforms; D012794:Sialic Acids / Q000378:metabolism; D012964:Sodium / Q000737:chemistry; D015222:Sodium Channels / Q000737:chemistry*; D013997:Time Factors; D014162:Transfection
D002241:Carbohydrates; D018076:DNA, Complementary; D020033:Protein Isoforms; D012794:Sialic Acids; D015222:Sodium Channels; D012964:Sodium; D019158:N-Acetylneuraminic Acid