TY - CHAP
T1 - Engineering K+ channels using semisynthesis
AU - Komarov, Alexander G.
AU - Costantino, Corey A.
AU - Valiyaveetil, Francis I.
N1 - Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
KW - K channels
KW - Membrane proteins
KW - Semisynthesis
KW - Solid-phase peptide synthesis
UR - http://www.scopus.com/inward/record.url?scp=84877048260&partnerID=8YFLogxK
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U2 - 10.1007/978-1-62703-345-9_1
DO - 10.1007/978-1-62703-345-9_1
M3 - Chapter
C2 - 23494368
AN - SCOPUS:84877048260
SN - 9781627033442
T3 - Methods in Molecular Biology
SP - 3
EP - 17
BT - Chemical Neurobiology
A2 - Banghart, Matthew
ER -