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Modification of Na channel inactivation by α-chymotrypsin in single cardiac myocytes

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Abstract

The effects of α-chymotrypsin and trypsin on the macroscopic Na current in isolated guinea pig ventricular myocytes at 16 ° C were investigated using the whole-cell voltage-clamp technique. Intracellular application of both enzymes reduced the extent of Na current inactivation during 20- to 50-ms depolarizing pulses. Elimination of fast inactivation by α-chymotrypsin was accompanied by a slowing of the rate of Na current decay through changes in both the time constants of current decay and the proportions of current undergoing a fast vs slow rate of decay. Treatment that reduced Na current decay to ⩽ 10 % within 20 ms was accompanied by a hyperpolarizing shift of the Na conductance/voltage relationship and an increase in the time-to-peak current that was most prominent for small depolarizations. Evidence for a significant slow inactivation process was obtained following removal of fast inactivation. The effect of trypsin (0.15–0.3 mg/ml) was less specific than α-chymotrypsin in that it also reduced Na conductance and increased leak current.

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References

  • Armstrong CM, Bezanilla F, Rojas E (1973) Destruction of sodium conductance inactivation in squid giant axons perfused with pronase. J Gen Physiol 62: 375–391

    Google Scholar 

  • Bezanilla F (1987) Single sodium channels from the squid giant axon. Biophys J 52: 1087–1090

    Google Scholar 

  • Carmeliet E (1987) Slow inactivation of the sodium current in rabbit cardiac Purkinje fibers. Pflügers Arch 408: 18–26

    Google Scholar 

  • Clarkson CW (1988) Effects of proteolytic enzymes on cardiac sodium channel inactivation and block by local anesthetics (abstract). Biophys J 53: 534a

    Google Scholar 

  • Clarkson CW, Matsubara T, Hondeghem LM (1984) Slow inactivation of V max in guinea pig ventricular myocardium. Am J Physiol 247: H645-H654

    Google Scholar 

  • Clarkson CW, Follmer CH, Ten Eick RE, Hondeghem LM, Yeh JZ (1988) Evidence for two components of sodium channel block by lidocaine in isolated cardiac myocytes. Circ Res 63: 869–878

    Google Scholar 

  • Cota G, Armstrong CM (1989) Sodium channel gating in clonal pituitary cells. The inactivation step is not voltage dependent. J Gen Physiol 94: 213–232

    Google Scholar 

  • Eaton DC, Brodwick MS, Oxford GS, Rudy B (1978) Arginine specific reagents remove sodium channel inactivation. Nature (Lond) 271: 473–476

    Google Scholar 

  • Fernandez JM, Fox AP, Krasne S (1984) Membrane patches and whole-cell membranes: a comparison of electrical properties in rat clonal pituitary (GH3) cells. J Physiol (Lond) 356: 565–585

    Google Scholar 

  • Follmer CF, Ten Eick RE, Yeh JZ (1987) Sodium current kinetics in cat atrial myocytes. J Physiol (Lond) 384: 169–197

    Google Scholar 

  • Fozzard HA, Hanck DA, Makielski JC, Scanley BE, Sheets MF (1987) Sodium channels in cardiac Purkinje cells. Experientia 43: 1162–1168

    Google Scholar 

  • Gintant GA, Datyner NB, Cohen IS (1984) Slow inactivation of a tetrodotoxin-sensitive current in canine cardiac Purkinje fibers. Biophys J 45: 509–512

    Google Scholar 

  • Gonoi T, Hille B (1987) Gating of sodium channels Inactivation modifiers discriminate between models. J Gen Physiol 89: 253–274

    Google Scholar 

  • Gordon D, Merrick D, Auld V, Dunn R, Goldin AL, Davidson N, Catterall WA (1987) Tissue-specific expression of the RI and RII sodium channel subtypes. Proc Natl Acad Sci USA 84: 8682–8686

    Google Scholar 

  • Guy HR (1988) A model relating the structure of the sodium channel to its function. Curr Top Membr Transp 33: 289–308

    Google Scholar 

  • Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 39: 85–100

    Google Scholar 

  • Huang JMC, Tanguy J, Yeh JZ (1987) Removal of sodium inactivation and block of sodium channels by chloramine-T in crayfish and squid giant axons. Biophys J 52: 155–163

    Google Scholar 

  • Makielski JC, Sheets MF, Hanck DA, January CT, Fozzard HA (1987) Sodium current in voltage clamped internally perfused canine cardiac Purkinje cells. Biophys J 52: 1–11

    Google Scholar 

  • Mitra R, Morad M (1985) A uniform enzymatic method for dissociation of myocytes from hearts and stomachs of vertebrates. Am J Physiol 249: H1056-H1060

    Google Scholar 

  • Neumcke B, Schwarz JR, Stampfli R (1987) A comparison os sodium currents in rat and frog myelinated nerve: normal and modified sodium inactivation. J Physiol (Lond) 382: 175–191

    Google Scholar 

  • Noda M, Ikeda T, Kayano T, Suzuki H, Takeshima H, Kurasaki M, Takahashi H, Numa S (1986) Existence of distinct sodium channel messenger RNAs in rat brain. Nature 320: 188–192

    Google Scholar 

  • Nonner W, Spalding BC, Hille B (1980) Low intracellular pH and chemical agents slow inactivation gating in sodium channels of muscle. Nature 284: 360–363

    Google Scholar 

  • Oxford GS, Wu CH, Narahashi T (1978) Removal of sodium channel inactivation in squid giant axons by N-bromoacetamide. J Gen Physiol 71: 227–247

    Google Scholar 

  • Rojas E, Rudy B (1976) Destruction of the sodium conductance inactivation by a specific protease in perfused nerve fibres from Loligo. J Physiol (Lond) 262: 501–531

    Google Scholar 

  • Rudy B (1978) Slow inactivation of the sodium conductance in squid giant axons. Pronase resistance. J Physiol (Lond) 283: 1–21

    Google Scholar 

  • Saikawa T, Carmeliet E (1982) Slow recovery of the maximal rate of rise (V max) of the action potential in sheep cardiac Purkinje fibers. Pflügers Arch 394: 90–93

    Google Scholar 

  • Vandenberg CA, Horn R (1984) Inactivation viewed through single sodium channels. J Gen Physiol 84: 535–564

    Google Scholar 

  • Vassilev PM, Scheuer T, Catterall WA (1988) Identification of an intracellular peptide segment involved in sodium channel inactivation. Science 241: 1658–1661

    Google Scholar 

  • Wang GK (1984a) Modification of sodium channel inactivation in single myelinated nerve fibers by methionine-reactive chemicals. Biophys J 46: 121–124

    Google Scholar 

  • Wang GK (1984b) Irreversible modification of sodium channel inactivation in toad myelinated nerve fibers by the oxidant chloramine-T. J Physiol (Lond) 346: 127–141

    Google Scholar 

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Clarkson, C.W. Modification of Na channel inactivation by α-chymotrypsin in single cardiac myocytes. Pflugers Arch. 417, 48–57 (1990). https://doi.org/10.1007/BF00370768

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  • DOI: https://doi.org/10.1007/BF00370768

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