Skip to main content
Log in

Effects of chemical modification of amino groups by two different imidoesters on voltage-clamped nerve fibres of the frog

  • Excitable Tissues and Central Nervous Physiology
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

Voltage clamped single nerve fibres of the frogPanta esculenta were treated with the amino groups specific reagents ethyl acetimidate and isethionyl acetimidate. Ethyl acetimidate is lipid soluble, relatively non-polar and can penetrate a membrane. Isethionyl acetimidate is lipid-insoluble, polar and membrane-impermeant. Treatment with ethyl acetimidate caused an irreversible reduction of Na currents and a shift of the voltage dependence of the steady-state sodium inactivation,h (E), in the hyperpolarizing direction. The voltage dependence of sodium activation was much less affected and a small shift into the depolarizing direction was observed. By contrast, the non-permeant reagent did not reduce the sodium currents and the shifts of theh (E) curve were smaller than the shifts caused by ethyl acetimidate. Furthermore, a small shift of the voltagedependence of activation in the hyperpolarizing direction was observed. As the modification procedure with imidoesters does not cause a change of net charge, the results cannot be explained by an effect of surface charge modification, rather, they seem to be due to a direct effect of amino group modification on the voltage dependence of inactivation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

TNBS:

Trinitrobenzenesulfonic acid

IAI:

isethionyl acetimidate

EAI:

ethyl acetimidate

MOPS:

morpholinopropanesulfonic acid

TEA:

tetraethylammonium

CHES:

(2-cyclohexylamino)-ethanesulfonic acid

References

  • Anderson GW, Zimmerman JE, Callahan FM (1964) The use of esters of N-hydroxysuccinimide in peptide synthesis. J Am Chem Soc 86:1839–1842

    Google Scholar 

  • Brodwick MS, Eaton DC (1982) Chemical modification of excitable membranes. Proteins in the nervous system: Structure and function. Alan R. Liss, New York, pp 51–72

    Google Scholar 

  • Cahalan MD, Pappone PA (1981) Chemical modification of sodium channel surface charges in frog skeletal muscle by trinitrobenzene sulphonic acid. J Physiol 321:127–139

    Google Scholar 

  • Courtney KR (1975) Mechanism of frequency-dependent inhibition of sodium currents in frog myelinated nerve by the lidocaine derivative GEA 968. J Pharmacol Exp Ther 195:225–236

    Google Scholar 

  • Courtney KR (1979) Extracellular pH selectively modulates recovery from sodium inactivation in frog myelinated nerve. Biophys J 28:363–368

    Google Scholar 

  • Drouin H, Neumcke B (1974) Specific and unspecific charges at the sodium channels of the nerve membrane. Pflügers Arch Eur J Physiol 351:207–229

    Google Scholar 

  • Frankenhaeuser B (1960) Quantitative description of sodium currents in myelinated nerve fibres of Xenopus laevis. J Physiol 151:491–501

    Google Scholar 

  • Hodgkin AL, Huxley AF (1952) A quantitative description of membrane current and its application to conduction and excitation in nerve. J Physiol 117:500–544

    Google Scholar 

  • Hunter MJ, Ludwig ML (1962) The reaction of imidoesters with proteins and related small molecules. J Am Chem Soc 84:3491–3504

    Google Scholar 

  • Nonner W (1969) A new voltage clamp method for Ranvier nodes. Pflügers Arch Eur J Physiol 309:176–192

    Google Scholar 

  • Okuyama T, Satake K (1960) On the preparation and properties of 2,4,6-trinitrophenyl-amino acids and-peptides. J Biochem 47:454–466

    Google Scholar 

  • Rack M, Hu S, Rubly N, Waschow C (1984) Effects of chemical modification of amino and sulfhydryl groups on the voltageclamped frog node of Ranvier. Pflügers Arch Eur J Physiol 400:403–408

    Google Scholar 

  • Rack M (1984) Amino group modification without destruction of charge affects sodium currents in frog nerve. Pflügers Arch Eur J Physiol 402:R36

    Google Scholar 

  • Stämpfli R (1969) Dissection of single nerve fibres and measurements of membrane potential changes of Ranvier nodes by means of the double air gap method. In: Passow H, Stämpfli R (eds) Laboratory techniques in membrane biophysics. Springer, Berlin Heidelberg New York, pp 157–166

    Google Scholar 

  • Stämpfli R, Hille B (1976) Electrophysiology of the peripheral myelinated nerve. In: Llinás R, Precht W (eds) Frog neurobiology. Springer, Berlin Heidelberg New York, pp 3–32

    Google Scholar 

  • Strichartz GR (1973) The inhibition of sodium currents in myelinated nerve by quaternary derivatives of lidocaine. J Gen Physiol 62:37–57

    Google Scholar 

  • Whiteley NM, Berg HC (1974) Amidination of the outer and inner surfaces of the human erythrocyte membrane. J Mol Biol 87:541–561

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rack, M. Effects of chemical modification of amino groups by two different imidoesters on voltage-clamped nerve fibres of the frog. Pflugers Arch. 404, 126–130 (1985). https://doi.org/10.1007/BF00585407

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00585407

Key words

Navigation