Two-dimensional probability density analysis of single channel currents from reconstituted acetylcholine receptors and sodium channels. 1990

B U Keller, and M S Montal, and R P Hartshorne, and M Montal
Department of Biology, University of California, San Diego, La Jolla 92093-0319.

Two-dimensional probability density analysis of single channel current recordings was applied to two purified channel proteins reconstituted in planar lipid bilayers: Torpedo acetylcholine receptors and voltage-sensitive sodium channels from rat brain. The information contained in the dynamic history of the gating process, i.e., the time sequence of opening and closing events was extracted from two-dimensional distributions of transitions between identifiable states. This approach allows one to identify kinetic models consistent with the observables. Gating of acetylcholine receptors expresses "memory" of the transition history: the receptor has two channel open (O) states; the residence time in each of them strongly depends on both the preceding open time and the intervening closed interval. Correspondingly, the residence time in the closed (C) states depends on both the preceding open time and the preceding closed time. This result confirms the scheme that considers, at least, two transition pathways between the open and closed states and extends the details of the model in that it defines that the short-lived open state is primarily entered from long-lived closed states while the long-lived open state is accessed mainly through short-lived closed states. Since ligand binding to the acetylcholine-binding sites is a reaction with channel closed states, we infer that the longest closed state (approximately 19 ms) is unliganded, the intermediate closed state (approximately 2 ms) is singly liganded and makes transitions to the short open state (approximately 0.5 ms) and the shortest closed state (approximately 0.4 ms) is doubly liganded and isomerizes to long open states (approximately 5 ms). This is the simplest interpretation consistent with available data. In contrast, sodium channels modified with batrachotoxin to eliminate inactivation show no correlation in the sequence of channel opening and closing events, i.e., have no memory of the transition history. This result is, therefore, consistent with any kinetic scheme that considers a single transition pathway between open and closed states, and confirms the C-C-O model previously inferred from one-dimensional distribution analysis. The strategy described should be of general validity in the analysis of single channel events from channel proteins in both natural and reconstituted membranes.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008051 Lipid Bilayers Layers of lipid molecules which are two molecules thick. Bilayer systems are frequently studied as models of biological membranes. Bilayers, Lipid,Bilayer, Lipid,Lipid Bilayer
D008954 Models, Biological Theoretical representations that simulate the behavior or activity of biological processes or diseases. For disease models in living animals, DISEASE MODELS, ANIMAL is available. Biological models include the use of mathematical equations, computers, and other electronic equipment. Biological Model,Biological Models,Model, Biological,Models, Biologic,Biologic Model,Biologic Models,Model, Biologic
D011336 Probability The study of chance processes or the relative frequency characterizing a chance process. Probabilities
D011950 Receptors, Cholinergic Cell surface proteins that bind acetylcholine with high affinity and trigger intracellular changes influencing the behavior of cells. Cholinergic receptors are divided into two major classes, muscarinic and nicotinic, based originally on their affinity for nicotine and muscarine. Each group is further subdivided based on pharmacology, location, mode of action, and/or molecular biology. ACh Receptor,Acetylcholine Receptor,Acetylcholine Receptors,Cholinergic Receptor,Cholinergic Receptors,Cholinoceptive Sites,Cholinoceptor,Cholinoceptors,Receptors, Acetylcholine,ACh Receptors,Receptors, ACh,Receptor, ACh,Receptor, Acetylcholine,Receptor, Cholinergic,Sites, Cholinoceptive
D001921 Brain The part of CENTRAL NERVOUS SYSTEM that is contained within the skull (CRANIUM). Arising from the NEURAL TUBE, the embryonic brain is comprised of three major parts including PROSENCEPHALON (the forebrain); MESENCEPHALON (the midbrain); and RHOMBENCEPHALON (the hindbrain). The developed brain consists of CEREBRUM; CEREBELLUM; and other structures in the BRAIN STEM. Encephalon
D004557 Electric Organ In about 250 species of electric fishes, modified muscle fibers forming disklike multinucleate plates arranged in stacks like batteries in series and embedded in a gelatinous matrix. A large torpedo ray may have half a million plates. Muscles in different parts of the body may be modified, i.e., the trunk and tail in the electric eel, the hyobranchial apparatus in the electric ray, and extrinsic eye muscles in the stargazers. Powerful electric organs emit pulses in brief bursts several times a second. They serve to stun prey and ward off predators. A large torpedo ray can produce of shock of more than 200 volts, capable of stunning a human. (Storer et al., General Zoology, 6th ed, p672) Electric Organs,Organ, Electric,Organs, Electric
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D014101 Torpedo A genus of the Torpedinidae family consisting of several species. Members of this family have powerful electric organs and are commonly called electric rays. Electric Rays,Torpedinidae,Rays, Electric
D015222 Sodium Channels Ion channels that specifically allow the passage of SODIUM ions. A variety of specific sodium channel subtypes are involved in serving specialized functions such as neuronal signaling, CARDIAC MUSCLE contraction, and KIDNEY function. Ion Channels, Sodium,Ion Channel, Sodium,Sodium Channel,Sodium Ion Channels,Channel, Sodium,Channel, Sodium Ion,Channels, Sodium,Channels, Sodium Ion,Sodium Ion Channel

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