Semisynthetic K+ channels show that the constricted conformation of the selectivity filter is not the C-type inactivated state

Prasanna K. Devaraneni, Alexander G. Komarov, Corey A. Costantino, Jordan J. Devereaux, Kimberly Matulef, Francis Valiyaveetil

Research output: Contribution to journalArticle

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Abstract

C-type inactivation of K+ channels plays a key role in modulating cellular excitability. During C-type inactivation, the selectivity filter of a K+ channel changes conformation from a conductive to a nonconductive state. Crystal structures of the KcsA channel determined at low K+ or in the open state revealed a constricted conformation of the selectivity filter, which was proposed to represent the C-type inactivated state. However, structural studies on other K+ channels do not support the constricted conformation as the C-type inactivated state. In this study, we address whether the constricted conformation of the selectivity filter is in fact the C-type inactivated state. The constricted conformation can be blocked by substituting the first conserved glycine in the selectivity filter with the unnatural amino acid D-Alanine. Protein semisynthesis was used to introduce D-Alanine into the selectivity filters of the KcsA channel and the voltage-gated K+ channel KvAP. For semisynthesis of the KvAP channel, we developed a modular approach in which chemical synthesis is limited to the selectivity filter whereas the rest of the protein is obtained by recombinant means. Using the semisynthetic KcsA and KvAP channels, we show that blocking the constricted conformation of the selectivity filter does not prevent inactivation, which suggests that the constricted conformation is not the C-type inactivated state.

Original languageEnglish (US)
Pages (from-to)15698-15703
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume110
Issue number39
DOIs
Publication statusPublished - Sep 24 2013

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