Engineering K+ channels using semisynthesis

Alexander G. Komarov, Corey A. Costantino, Francis I. Valiyaveetil

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Potassium channels conduct K+ ions selectively and at very high rates. Central to the function of K+ channels is a structural unit called the selectivity filter. In the selectivity filter, a row of four K + binding sites are created using mainly the backbone carbonyl oxygen atoms. Due to the involvement of the protein backbone, site-directed mutagenesis is of limited utility in investigating the selectivity filter. In order to overcome this limitation, we have developed a semisynthetic approach, which permits the use of chemical synthesis to manipulate the selectivity filter. In this chapter, we describe the protocols that we have developed for the semisynthesis of the K+ channel, KcsA. We anticipate that the protocols described in this chapter will also be applicable for the semisynthesis of other integral membrane proteins of interest.

Original languageEnglish (US)
Title of host publicationChemical Neurobiology
Subtitle of host publicationMethods and Protocols
EditorsMatthew Banghart
Pages3-17
Number of pages15
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume995
ISSN (Print)1064-3745

Keywords

  • K channels
  • Membrane proteins
  • Semisynthesis
  • Solid-phase peptide synthesis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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