Isolation and characterization of a high affinity peptide inhibitor of ClC-2 chloride channels

Christopher H. Thompson, Pedro R. Olivetti, Matthew D. Fuller, Cody S. Freeman, Denis McMaster, Robert J. French, Jan Pohl, Julia Kubanek, Nael A. MaCarty

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

The ClC protein family includes voltage-gated chloride channels and chloride/proton exchangers. In eukaryotes, ClC proteins regulate membrane potential of excitable cells, contribute to epithelial transport, and aid in lysosomal acidification. Although structure/ function studies of ClC proteins have been aided greatly by the available crystal structures of a bacterial ClC chloride/proton exchanger, the availability of useful pharmacological tools, such as peptide toxin inhibitors, has lagged far behind that of their cation channel counterparts. Here we report the isolation, from Leiurus quinquestriatus hebraeus venom, of a peptide toxin inhibitor of the ClC-2 chloride channel. This toxin, GaTx2, inhibits ClC-2 channels with a voltage-dependent apparent KD of ∼20 pM, making it the highest affinity inhibitor of any chloride channel. GaTx2 slows ClC-2 activation by increasing the latency to first opening by nearly 8-fold but is unable to inhibit open channels, suggesting that this toxin inhibits channel activation gating. Finally, GaTx2 specifically inhibits ClC-2 channels, showing no inhibitory effect on a battery of other major classes of chloride channels and voltage-gated potassium channels. GaTx2 is the first peptide toxin inhibitor of any ClC protein. The high affinity and specificity displayed by this toxin will make it a very powerful pharmacological tool to probe ClC-2 structure/function.

Original languageEnglish (US)
Pages (from-to)26051-26052
Number of pages2
JournalJournal of Biological Chemistry
Volume284
Issue number38
DOIs
StatePublished - Sep 18 2009

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Isolation and characterization of a high affinity peptide inhibitor of ClC-2 chloride channels'. Together they form a unique fingerprint.

  • Cite this

    Thompson, C. H., Olivetti, P. R., Fuller, M. D., Freeman, C. S., McMaster, D., French, R. J., Pohl, J., Kubanek, J., & MaCarty, N. A. (2009). Isolation and characterization of a high affinity peptide inhibitor of ClC-2 chloride channels. Journal of Biological Chemistry, 284(38), 26051-26052. https://doi.org/10.1074/jbc.M109.031724