Abstract
Analysis of data from single-channel studies can provide us with insights into the detailed physicochemical features of channel proteins, such as ion permeation rate, ion selectivity, and gating. However, single-channel analysis, particularly gating analysis, is sometimes time-consuming, unconvincing, and unproductive. Notwithstanding, if it is combined with structure information and site-directed mutagenesis of channel proteins, it can prove to be a powerful tool that moves us toward the ultimate understanding of structure–function of channel proteins. This chapter is composed of roughly two parts. The first part describes the technical aspect how to set up the hardware for correct acquisition of the tiny, long-lasting single-channel currents against background noises and drifts. The second part deals with the statistical estimation of the current amplitude and dwell times of the open and closed states and how to fit the estimated data to an appropriate reaction model. Each section is described in a step-by-step fashion so even beginners can understand and experience the course of single-channel study, from data acquisition to model fitting.
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Notes
- 1.
The description on the data recording in this section seems to be rather out of date given the huge capacity of recent hard disks. However, this bygone description is left for the sake of researchers that use old computers due to A/D converters and software.
References
Miller C (ed) (1986) Ion channel reconstitution. Plenum, New York
Sakmann B, Neher E (eds) (1995) Single-channel recording, 2nd edn. New York, Plenum
Patlak JB (1988) Sodium channel subconductance levels measured with a new variance-mean analysis. J Gen Physiol 92:413–430
Hille B (1992) Ionic channels of excitable membranes, 2nd edn. Sinauer, Sunderland
Colquhoun D, Hawkes AG (1983) The principles of the stochastic interpretation of ion-channel mechanisms. In: Sakmann B, Neher E (eds) Single-channel recording. Plenum, New York, pp 135–189
Sachs F, Neil J, Barkakati N (1983) The automated analysis of data from single ionic channels. Pflugers Arch 395:331–340
Dempster J (1993) Computer analysis of electrophysiological signals. Academic Press, London, p167
Colquhoun D (1987) Practical analysis of single channel records. In: Standen NB, Gray PTA, Whitaker MJ (eds) Microelectrode techniques. The Plymus workshop handbook. Company of Biologist Ltd, Cambridge, pp 83–104
Sigworth FJ, Sine SM (1987) Data transformations for improved display and fitting of single channel dwell time histograms. Biophys J 48:149–158
Jackson MB (1992) Stationary single-channel analysis. Method Enzymol 207:729–746
Nakagawa T, Koyanagi Y (1982) Analysis of experimental data by least square method (in Japanese). University of Tokyo Press, Tokyo
Awaya T (1991) Data analysis (2nd edition, in Japanese). Japan Scientific Societies Press, Tokyo
French RJ, Wonderli WF (1992) Software for acquision and analysis of ion channel data:choices, tasks, and strategies. Method Enzymol 207:711–728
Colquhoun D, Hawkes AG (1981) On the stochastic properties of single ion channels. Proc R Soc Lond B 211:205–235
Colquhoun D, Hawkes AG (1983) On the stochastic properties of bursts of single ion channel openings and clusters of bursts. Phil Trans R Soc Lond B 300:1–59
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Sokabe, M. (2012). Methods for Processing and Analyzing Single-Channel Data. In: Okada, Y. (eds) Patch Clamp Techniques. Springer Protocols Handbooks. Springer, Tokyo. https://doi.org/10.1007/978-4-431-53993-3_5
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DOI: https://doi.org/10.1007/978-4-431-53993-3_5
Publisher Name: Springer, Tokyo
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