Interaction of local anesthetics with a peptide encompassing the IV/S4-S5 linker of the Na+ channel

Leonardo F. Fraceto, Sérgio Oyama, Clóvis R. Nakaie, Alberto Spisni, Eneida de Paula, Thelma A. Pertinhez

Research output: Contribution to journalArticle

Abstract

The peptide pIV/S4-S5 encompasses the cytoplasmic linker between helices S4-S5 in domain IV of the voltage-gated Na+ channel, residues 1644-1664.22Residues numbering correspond to the primary sequence of the voltage-gated human brain Na+ channel [1]. The interaction of two local anesthetics (LA), lidocaine and benzocaine, with pIV/S4-S5 has been studied by DOSY, heteronuclear NMR 1H-15N-HSQC spectroscopy and computational methods. DOSY indicates that benzocaine, a neutral ester, exhibits stronger interaction with pIV/S4-S5 than lidocaine, a charged amine-amide. Weighted average chemical shifts, Δδ(1H-15N), show that benzocaine affects residues L1653, M1655 and S1656 while lidocaine slightly perturbs residues I1646, L1649 and A1659, L1660, near the N- and C-terminus, respectively. Computational methods confirmed the stability of the benzocaine binding and the existence of two binding sites for lidocaine. Even considering that the approach of studying the peptide in the presence of a co-solvent (TFE/H2O, 30%/70% v/v) has an inherently limited implication, our data strongly support the existence of multiple LA binding sites in the IV/S4-S5 linker, as suggested in the literature. In addition, we consider that LA can bind to the S4-S5 linker with diverse binding modes and strength since this linker is part of the receptor for the "inactivation gate particle". Conditions for devising new functional studies, aiming to better understand Na+ channel functionality as well as the various facets of LA pharmacological activity are proposed in this work.

Original languageEnglish
Pages (from-to)29-39
Number of pages11
JournalBiophysical Chemistry
Volume123
Issue number1
DOIs
Publication statusPublished - Aug 20 2006

Fingerprint

Benzocaine
anesthetics
Lidocaine
Local Anesthetics
peptides
Peptides
Computational methods
Binding Sites
interactions
Biomolecular Nuclear Magnetic Resonance
Polytetrafluoroethylene
Chemical shift
Electric potential
electric potential
activity (biology)
Amides
deactivation
helices
amides
Amines

Keywords

  • Benzocaine
  • Lidocaine
  • Local anesthetics
  • Nuclear magnetic resonance
  • Peptide
  • Voltage-gated sodium channel

ASJC Scopus subject areas

  • Biochemistry
  • Physical and Theoretical Chemistry
  • Biophysics

Cite this

Fraceto, L. F., Oyama, S., Nakaie, C. R., Spisni, A., de Paula, E., & Pertinhez, T. A. (2006). Interaction of local anesthetics with a peptide encompassing the IV/S4-S5 linker of the Na+ channel. Biophysical Chemistry, 123(1), 29-39. https://doi.org/10.1016/j.bpc.2006.03.010

Interaction of local anesthetics with a peptide encompassing the IV/S4-S5 linker of the Na+ channel. / Fraceto, Leonardo F.; Oyama, Sérgio; Nakaie, Clóvis R.; Spisni, Alberto; de Paula, Eneida; Pertinhez, Thelma A.

In: Biophysical Chemistry, Vol. 123, No. 1, 20.08.2006, p. 29-39.

Research output: Contribution to journalArticle

Fraceto, LF, Oyama, S, Nakaie, CR, Spisni, A, de Paula, E & Pertinhez, TA 2006, 'Interaction of local anesthetics with a peptide encompassing the IV/S4-S5 linker of the Na+ channel', Biophysical Chemistry, vol. 123, no. 1, pp. 29-39. https://doi.org/10.1016/j.bpc.2006.03.010
Fraceto, Leonardo F. ; Oyama, Sérgio ; Nakaie, Clóvis R. ; Spisni, Alberto ; de Paula, Eneida ; Pertinhez, Thelma A. / Interaction of local anesthetics with a peptide encompassing the IV/S4-S5 linker of the Na+ channel. In: Biophysical Chemistry. 2006 ; Vol. 123, No. 1. pp. 29-39.
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abstract = "The peptide pIV/S4-S5 encompasses the cytoplasmic linker between helices S4-S5 in domain IV of the voltage-gated Na+ channel, residues 1644-1664.22Residues numbering correspond to the primary sequence of the voltage-gated human brain Na+ channel [1]. The interaction of two local anesthetics (LA), lidocaine and benzocaine, with pIV/S4-S5 has been studied by DOSY, heteronuclear NMR 1H-15N-HSQC spectroscopy and computational methods. DOSY indicates that benzocaine, a neutral ester, exhibits stronger interaction with pIV/S4-S5 than lidocaine, a charged amine-amide. Weighted average chemical shifts, Δδ(1H-15N), show that benzocaine affects residues L1653, M1655 and S1656 while lidocaine slightly perturbs residues I1646, L1649 and A1659, L1660, near the N- and C-terminus, respectively. Computational methods confirmed the stability of the benzocaine binding and the existence of two binding sites for lidocaine. Even considering that the approach of studying the peptide in the presence of a co-solvent (TFE/H2O, 30{\%}/70{\%} v/v) has an inherently limited implication, our data strongly support the existence of multiple LA binding sites in the IV/S4-S5 linker, as suggested in the literature. In addition, we consider that LA can bind to the S4-S5 linker with diverse binding modes and strength since this linker is part of the receptor for the {"}inactivation gate particle{"}. Conditions for devising new functional studies, aiming to better understand Na+ channel functionality as well as the various facets of LA pharmacological activity are proposed in this work.",
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AU - Fraceto, Leonardo F.

AU - Oyama, Sérgio

AU - Nakaie, Clóvis R.

AU - Spisni, Alberto

AU - de Paula, Eneida

AU - Pertinhez, Thelma A.

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N2 - The peptide pIV/S4-S5 encompasses the cytoplasmic linker between helices S4-S5 in domain IV of the voltage-gated Na+ channel, residues 1644-1664.22Residues numbering correspond to the primary sequence of the voltage-gated human brain Na+ channel [1]. The interaction of two local anesthetics (LA), lidocaine and benzocaine, with pIV/S4-S5 has been studied by DOSY, heteronuclear NMR 1H-15N-HSQC spectroscopy and computational methods. DOSY indicates that benzocaine, a neutral ester, exhibits stronger interaction with pIV/S4-S5 than lidocaine, a charged amine-amide. Weighted average chemical shifts, Δδ(1H-15N), show that benzocaine affects residues L1653, M1655 and S1656 while lidocaine slightly perturbs residues I1646, L1649 and A1659, L1660, near the N- and C-terminus, respectively. Computational methods confirmed the stability of the benzocaine binding and the existence of two binding sites for lidocaine. Even considering that the approach of studying the peptide in the presence of a co-solvent (TFE/H2O, 30%/70% v/v) has an inherently limited implication, our data strongly support the existence of multiple LA binding sites in the IV/S4-S5 linker, as suggested in the literature. In addition, we consider that LA can bind to the S4-S5 linker with diverse binding modes and strength since this linker is part of the receptor for the "inactivation gate particle". Conditions for devising new functional studies, aiming to better understand Na+ channel functionality as well as the various facets of LA pharmacological activity are proposed in this work.

AB - The peptide pIV/S4-S5 encompasses the cytoplasmic linker between helices S4-S5 in domain IV of the voltage-gated Na+ channel, residues 1644-1664.22Residues numbering correspond to the primary sequence of the voltage-gated human brain Na+ channel [1]. The interaction of two local anesthetics (LA), lidocaine and benzocaine, with pIV/S4-S5 has been studied by DOSY, heteronuclear NMR 1H-15N-HSQC spectroscopy and computational methods. DOSY indicates that benzocaine, a neutral ester, exhibits stronger interaction with pIV/S4-S5 than lidocaine, a charged amine-amide. Weighted average chemical shifts, Δδ(1H-15N), show that benzocaine affects residues L1653, M1655 and S1656 while lidocaine slightly perturbs residues I1646, L1649 and A1659, L1660, near the N- and C-terminus, respectively. Computational methods confirmed the stability of the benzocaine binding and the existence of two binding sites for lidocaine. Even considering that the approach of studying the peptide in the presence of a co-solvent (TFE/H2O, 30%/70% v/v) has an inherently limited implication, our data strongly support the existence of multiple LA binding sites in the IV/S4-S5 linker, as suggested in the literature. In addition, we consider that LA can bind to the S4-S5 linker with diverse binding modes and strength since this linker is part of the receptor for the "inactivation gate particle". Conditions for devising new functional studies, aiming to better understand Na+ channel functionality as well as the various facets of LA pharmacological activity are proposed in this work.

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