Conformational dynamics in the selectivity filter of KcsA in response to potassium ion concentration

Manasi P. Bhate, Benjamin J. Wylie, Lin Tian, Ann E. McDermott

Research output: Contribution to journalArticlepeer-review

67 Scopus citations

Abstract

Conformational change in the selectivity filter of KcsA as a function of ambient potassium concentration is studied with solid-state NMR. This highly conserved region of the protein is known to chelate potassium ions selectively. We report solid-state NMR chemical shift fingerprints of two distinct conformations of the selectivity filter; significant changes are observed in the chemical shifts of key residues in the filter as the potassium ion concentration is changed from 50 mM to 1 μM. Potassium ion titration studies reveal that the site-specific Kd for K+ binding at the key pore residue Val76 is on the order of ~7 μM and that a relatively high sample hydration is necessary to observe the low-K+ conformer. Simultaneous detection of both conformers at low ambient potassium concentration suggests that the high-K+ and low-K+ states are in slow exchange on the NMR timescale (kex<500 s-1). The slow rate and tight binding for evacuating both inner sites simultaneously differ from prior observations in detergent in solution, but agree well with measurements by electrophysiology and appear to result from our use of a hydrated bilayer environment. These observations strongly support a common assumption that the low-K+ state is not involved in ion transmission, and that during transmission one of the two inner sites is always occupied. On the other hand, these kinetic and thermodynamic characteristics of the evacuation of the inner sites certainly could be compatible with participation in a control mechanism at low ion concentration such as C-type inactivation, a process that is coupled to activation and involves closing of the outer mouth of the channel.

Original languageEnglish
Pages (from-to)155-166
Number of pages12
JournalJournal of molecular biology
Volume401
Issue number2
DOIs
StatePublished - Aug 2010

Keywords

  • Conformational dynamics
  • KcsA
  • Potassium channel
  • Selectivity filter
  • Solid-state NMR

Fingerprint Dive into the research topics of 'Conformational dynamics in the selectivity filter of KcsA in response to potassium ion concentration'. Together they form a unique fingerprint.

Cite this