The interaction between delayed rectifier channel alpha-subunits does not involve hetero-tetramer formation

P. Biliczki, Andre Rüdiger, Zenawit Girmatsion, Marc Pourrier, Aida M. Mamarbachi, Terence E. Hébert, Ralf P. Brandes, Stefan H. Hohnloser, Stanley Nattel, Joachim R. Ehrlich

Research output: Contribution to journalArticle

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Abstract

Abstract We have previously reported a physiologically relevant interaction between KCNQ1 (Q1) and KCNH2 (H2). While the H2 C-terminus has been suggested to play a role, so far, no more detailed information regarding the interaction site is available. The methods used in the study are cell culture, PCR for mutagenesis, patch clamp for ion current recordings, co-immunoprecipitation for determination of protein interaction. Co-expression of Q1 and H2 resulted in an increase of I H2 (tails after +50 mV; Q1+H2, 36±6 pA/pF; H2, 14±2 pA/pF; n=10; 12; PH2 (tails after +50 mV; H2+dnQ1, 24±4 pA/pF; n=10; P50). However, I H2 sensitivity did not significantly change in the presence or absence of Q1 (IC50 341±63 vs. 611±293 nmol/L, respectively, P=n.s.), providing further evidence that the pore is not a likely H2-Q1 interaction site. To obtain further insights into the role of intra-cytoplasmic structures, we used both C- and N-terminally truncated mutant H2 proteins. Both H2 mutants co-immunoprecipitated with Q1, suggesting no specific role of C- or N-termini. Accordingly, rather than these, the transmembrane domains of the α-subunits appear relevant for the interaction. Our results largely exclude the formation of hetero-tetramers between H2 and Q1 comprising the pore region or H2 C- or N-termini.

Original languageEnglish
Article number1108
Pages (from-to)973-981
Number of pages9
JournalNaunyn-Schmiedeberg's Archives of Pharmacology
Volume388
Issue number9
DOIs
Publication statusPublished - Sep 1 2015

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Cytoplasmic Structures
Mutant Proteins
Immunoprecipitation
Mutagenesis
Inhibitory Concentration 50
Cell Culture Techniques
Ions
Polymerase Chain Reaction
Proteins

Keywords

  • Hetero-tetramer
  • KCNH2
  • KCNQ1
  • protein interaction

ASJC Scopus subject areas

  • Pharmacology

Cite this

The interaction between delayed rectifier channel alpha-subunits does not involve hetero-tetramer formation. / Biliczki, P.; Rüdiger, Andre; Girmatsion, Zenawit; Pourrier, Marc; Mamarbachi, Aida M.; Hébert, Terence E.; Brandes, Ralf P.; Hohnloser, Stefan H.; Nattel, Stanley; Ehrlich, Joachim R.

In: Naunyn-Schmiedeberg's Archives of Pharmacology, Vol. 388, No. 9, 1108, 01.09.2015, p. 973-981.

Research output: Contribution to journalArticle

Biliczki, P, Rüdiger, A, Girmatsion, Z, Pourrier, M, Mamarbachi, AM, Hébert, TE, Brandes, RP, Hohnloser, SH, Nattel, S & Ehrlich, JR 2015, 'The interaction between delayed rectifier channel alpha-subunits does not involve hetero-tetramer formation', Naunyn-Schmiedeberg's Archives of Pharmacology, vol. 388, no. 9, 1108, pp. 973-981. https://doi.org/10.1007/s00210-015-1108-3
Biliczki, P. ; Rüdiger, Andre ; Girmatsion, Zenawit ; Pourrier, Marc ; Mamarbachi, Aida M. ; Hébert, Terence E. ; Brandes, Ralf P. ; Hohnloser, Stefan H. ; Nattel, Stanley ; Ehrlich, Joachim R. / The interaction between delayed rectifier channel alpha-subunits does not involve hetero-tetramer formation. In: Naunyn-Schmiedeberg's Archives of Pharmacology. 2015 ; Vol. 388, No. 9. pp. 973-981.
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abstract = "Abstract We have previously reported a physiologically relevant interaction between KCNQ1 (Q1) and KCNH2 (H2). While the H2 C-terminus has been suggested to play a role, so far, no more detailed information regarding the interaction site is available. The methods used in the study are cell culture, PCR for mutagenesis, patch clamp for ion current recordings, co-immunoprecipitation for determination of protein interaction. Co-expression of Q1 and H2 resulted in an increase of I H2 (tails after +50 mV; Q1+H2, 36±6 pA/pF; H2, 14±2 pA/pF; n=10; 12; PH2 (tails after +50 mV; H2+dnQ1, 24±4 pA/pF; n=10; P50). However, I H2 sensitivity did not significantly change in the presence or absence of Q1 (IC50 341±63 vs. 611±293 nmol/L, respectively, P=n.s.), providing further evidence that the pore is not a likely H2-Q1 interaction site. To obtain further insights into the role of intra-cytoplasmic structures, we used both C- and N-terminally truncated mutant H2 proteins. Both H2 mutants co-immunoprecipitated with Q1, suggesting no specific role of C- or N-termini. Accordingly, rather than these, the transmembrane domains of the α-subunits appear relevant for the interaction. Our results largely exclude the formation of hetero-tetramers between H2 and Q1 comprising the pore region or H2 C- or N-termini.",
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AU - Rüdiger, Andre

AU - Girmatsion, Zenawit

AU - Pourrier, Marc

AU - Mamarbachi, Aida M.

AU - Hébert, Terence E.

AU - Brandes, Ralf P.

AU - Hohnloser, Stefan H.

AU - Nattel, Stanley

AU - Ehrlich, Joachim R.

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N2 - Abstract We have previously reported a physiologically relevant interaction between KCNQ1 (Q1) and KCNH2 (H2). While the H2 C-terminus has been suggested to play a role, so far, no more detailed information regarding the interaction site is available. The methods used in the study are cell culture, PCR for mutagenesis, patch clamp for ion current recordings, co-immunoprecipitation for determination of protein interaction. Co-expression of Q1 and H2 resulted in an increase of I H2 (tails after +50 mV; Q1+H2, 36±6 pA/pF; H2, 14±2 pA/pF; n=10; 12; PH2 (tails after +50 mV; H2+dnQ1, 24±4 pA/pF; n=10; P50). However, I H2 sensitivity did not significantly change in the presence or absence of Q1 (IC50 341±63 vs. 611±293 nmol/L, respectively, P=n.s.), providing further evidence that the pore is not a likely H2-Q1 interaction site. To obtain further insights into the role of intra-cytoplasmic structures, we used both C- and N-terminally truncated mutant H2 proteins. Both H2 mutants co-immunoprecipitated with Q1, suggesting no specific role of C- or N-termini. Accordingly, rather than these, the transmembrane domains of the α-subunits appear relevant for the interaction. Our results largely exclude the formation of hetero-tetramers between H2 and Q1 comprising the pore region or H2 C- or N-termini.

AB - Abstract We have previously reported a physiologically relevant interaction between KCNQ1 (Q1) and KCNH2 (H2). While the H2 C-terminus has been suggested to play a role, so far, no more detailed information regarding the interaction site is available. The methods used in the study are cell culture, PCR for mutagenesis, patch clamp for ion current recordings, co-immunoprecipitation for determination of protein interaction. Co-expression of Q1 and H2 resulted in an increase of I H2 (tails after +50 mV; Q1+H2, 36±6 pA/pF; H2, 14±2 pA/pF; n=10; 12; PH2 (tails after +50 mV; H2+dnQ1, 24±4 pA/pF; n=10; P50). However, I H2 sensitivity did not significantly change in the presence or absence of Q1 (IC50 341±63 vs. 611±293 nmol/L, respectively, P=n.s.), providing further evidence that the pore is not a likely H2-Q1 interaction site. To obtain further insights into the role of intra-cytoplasmic structures, we used both C- and N-terminally truncated mutant H2 proteins. Both H2 mutants co-immunoprecipitated with Q1, suggesting no specific role of C- or N-termini. Accordingly, rather than these, the transmembrane domains of the α-subunits appear relevant for the interaction. Our results largely exclude the formation of hetero-tetramers between H2 and Q1 comprising the pore region or H2 C- or N-termini.

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KW - KCNQ1

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