Local and global structure of the monomeric subunit of the potassium channel KcsA probed by NMR

• Published in: Biochimica et biophysica acta Vol. 1768; no. 12; pp. 3260 - 3270
• Main Authors: Chill, Jordan H., Louis, John M., Delaglio, Frank, Bax, Ad
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2007
• Subjects: Circular Dichroism; Magnetic Resonance Spectroscopy - methods; Membrane proteins; Models, Biological; NMR; Potassium channel; Potassium Channels - chemistry; Protein Subunits; Protons; RDC; Solvent exchange; RDC; NMR; Solvent exchange; Potassium channel; Membrane proteins
• ISSN: 0005-2736; 0006-3002; 1879-2642
• DOI: 10.1016/j.bbamem.2007.08.006

KcsA is a homotetrameric 68-kDa membrane-associated potassium channel which selectively gates the flux of potassium ions across the membrane. The channel is known to undergo a pH-dependent open-to-closed transition. Here we describe an NMR study of the monomeric subunit of the channel (KcsA M), solubilized in SDS micelles. Chemical shift, solvent exchange, backbone 15N relaxation and residual dipolar coupling (RDC) data show the TM1 helix to remain intact, but the TM2 helix contains a distinct kink, which is subject to concentration-independent but pH-dependent conformational exchange on a microsecond time scale. The kink region, centered at G99, was previously implicated in the gating of the tetrameric KcsA channel. An RDC-based model of KcsA M at acidic pH orients TM1 and the two helical segments of the kinked TM2 in a configuration reminiscent of the open conformation of the channel. Thus, the transition between states appears to be an inherent capability of the monomer, with the tetrameric assembly exerting a modulatory effect upon the transition which gives the channel its physiological gating profile.