Voltage-gated K+ channels contain multiple intersubunit association sites

LiWei Tu, Vincent Santarelli, ZuFang Sheng, William Skach, Debkumar Pain, Carol Deutsch

Research output: Contribution to journalArticle

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Abstract

A domain in the cytoplasmic NH2 terminus of voltage-gated K+ channels supervises the proper assembly of specific tetrameric channels (Li, M., Jan, J. M., and Jan, L. Y. (1992) Science 257, 1225-1230; Shen, N. V., Chen X., Boyer, M. M., and Pfaffinger, P. (1993) Neuron 11, 67-76). It is referred to as a first tetramerization domain, or T1 (Shen, N. V., Chen X., Boyer, M. M., and Pfaffinger, P. (1993) Neuron 11, 67-76). However, a deletion mutant of Kv1.3 that lacks the first 141 amino acids, Kv1.3 (T1-) forms functional channels, suggesting that additional association sites in the central core of Kv1.3 mediate oligomerization. To characterize these sites, we have tested the abilities of cRNA Kv1.3 (T1-) fragments coinjected with Kv1.3 (T1-) to suppress current in Xenopus oocytes. The fragments include portions of the six putative transmembrane segments, S1 through S6, specifically: S1, S1-S2, S1-S2-S3, S2-S3, S2-S3-S4, S3-S4, S3-S4-S5, S2 through COOH, S3 through COOH, S4 through COOH, and S5-S6-COOH. Electrophysiologic experiments show that the fragments S1-S2-S3, S3-S4-S5, S2 through COOH, and S3 through COOH strongly suppress Kv1.3 (T1-) current, while others do not. Suppression of expressed current is due to specific effects of the translated peptide Kv1.3 fragments, as validated by in vivo immunoprecipitation studies of a strong suppressor and a nonsuppressor. Pulse-chase experiments indicate that translation of truncated peptide fragments neither prevents translation of Kv1.3 (T1-) nor increases its rate of degradation. Co-immunoprecipitation experiments suggest that suppression involves direct association of a peptide fragment with Kv1.3 (T1-). Fragments that strongly suppress Kv1.3 (T1-) also suppress an analogous NH2-terminal deletion mutant of Kv2.1 (Kv2.1 (ΔN139)), an isoform belonging to a different subfamily. Our results indicate that sites in the central core of Kv1.3 facilitate intersubunit association and that there are suppression sites in the central core, which are promiscuous across voltage- gated K+ channel subfamilies.

Original languageEnglish (US)
Pages (from-to)18904-18911
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number31
DOIs
StatePublished - 1996
Externally publishedYes

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Voltage-Gated Potassium Channels
Peptide Fragments
S 6
Association reactions
Immunoprecipitation
Neurons
Complementary RNA
Oligomerization
Experiments
Xenopus
Oocytes
Protein Isoforms
Amino Acids
Degradation
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tu, L., Santarelli, V., Sheng, Z., Skach, W., Pain, D., & Deutsch, C. (1996). Voltage-gated K+ channels contain multiple intersubunit association sites. Journal of Biological Chemistry, 271(31), 18904-18911. https://doi.org/10.1074/jbc.271.31.18904

Voltage-gated K+ channels contain multiple intersubunit association sites. / Tu, LiWei; Santarelli, Vincent; Sheng, ZuFang; Skach, William; Pain, Debkumar; Deutsch, Carol.

In: Journal of Biological Chemistry, Vol. 271, No. 31, 1996, p. 18904-18911.

Research output: Contribution to journalArticle

Tu, L, Santarelli, V, Sheng, Z, Skach, W, Pain, D & Deutsch, C 1996, 'Voltage-gated K+ channels contain multiple intersubunit association sites', Journal of Biological Chemistry, vol. 271, no. 31, pp. 18904-18911. https://doi.org/10.1074/jbc.271.31.18904
Tu, LiWei ; Santarelli, Vincent ; Sheng, ZuFang ; Skach, William ; Pain, Debkumar ; Deutsch, Carol. / Voltage-gated K+ channels contain multiple intersubunit association sites. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 31. pp. 18904-18911.
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abstract = "A domain in the cytoplasmic NH2 terminus of voltage-gated K+ channels supervises the proper assembly of specific tetrameric channels (Li, M., Jan, J. M., and Jan, L. Y. (1992) Science 257, 1225-1230; Shen, N. V., Chen X., Boyer, M. M., and Pfaffinger, P. (1993) Neuron 11, 67-76). It is referred to as a first tetramerization domain, or T1 (Shen, N. V., Chen X., Boyer, M. M., and Pfaffinger, P. (1993) Neuron 11, 67-76). However, a deletion mutant of Kv1.3 that lacks the first 141 amino acids, Kv1.3 (T1-) forms functional channels, suggesting that additional association sites in the central core of Kv1.3 mediate oligomerization. To characterize these sites, we have tested the abilities of cRNA Kv1.3 (T1-) fragments coinjected with Kv1.3 (T1-) to suppress current in Xenopus oocytes. The fragments include portions of the six putative transmembrane segments, S1 through S6, specifically: S1, S1-S2, S1-S2-S3, S2-S3, S2-S3-S4, S3-S4, S3-S4-S5, S2 through COOH, S3 through COOH, S4 through COOH, and S5-S6-COOH. Electrophysiologic experiments show that the fragments S1-S2-S3, S3-S4-S5, S2 through COOH, and S3 through COOH strongly suppress Kv1.3 (T1-) current, while others do not. Suppression of expressed current is due to specific effects of the translated peptide Kv1.3 fragments, as validated by in vivo immunoprecipitation studies of a strong suppressor and a nonsuppressor. Pulse-chase experiments indicate that translation of truncated peptide fragments neither prevents translation of Kv1.3 (T1-) nor increases its rate of degradation. Co-immunoprecipitation experiments suggest that suppression involves direct association of a peptide fragment with Kv1.3 (T1-). Fragments that strongly suppress Kv1.3 (T1-) also suppress an analogous NH2-terminal deletion mutant of Kv2.1 (Kv2.1 (ΔN139)), an isoform belonging to a different subfamily. Our results indicate that sites in the central core of Kv1.3 facilitate intersubunit association and that there are suppression sites in the central core, which are promiscuous across voltage- gated K+ channel subfamilies.",
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AU - Santarelli, Vincent

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AU - Deutsch, Carol

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