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Title: Structural and Thermodynamic Properties of Selective Ion Binding in a K+ Channel

Abstract

Thermodynamic measurements of ion binding to the Streptomyces lividans K+ channel were carried out using isothermal titration calorimetry, whereas atomic structures of ion-bound and ion-free conformations of the channel were characterized by x-ray crystallography. Here we use these assays to show that the ion radius dependence of selectivity stems from the channel's recognition of ion size (i.e., volume) rather than charge density. Ion size recognition is a function of the channel's ability to adopt a very specific conductive structure with larger ions (K+, Rb+, Cs+, and Ba2+) bound and not with smaller ions (Na+, Mg2+, and Ca2+). The formation of the conductive structure involves selectivity filter atoms that are in direct contact with bound ions as well as protein atoms surrounding the selectivity filter up to a distance of 15 Angstroms from the ions. We conclude that ion selectivity in a K+ channel is a property of size-matched ion binding sites created by the protein structure.

Authors:
; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
960132
Report Number(s):
BNL-83118-2009-JA
TRN: US201016%%1276
DOE Contract Number:
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: PLoS Biology; Journal Volume: 5; Journal Issue: 5
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; ATOMS; CALORIMETRY; CHARGE DENSITY; CRYSTALLOGRAPHY; PROTEIN STRUCTURE; PROTEINS; STREPTOMYCES; THERMODYNAMIC PROPERTIES; THERMODYNAMICS; TITRATION; national synchrotron light source

Citation Formats

Lockless,S., Zhou, M., and MacKinnon, R.. Structural and Thermodynamic Properties of Selective Ion Binding in a K+ Channel. United States: N. p., 2007. Web. doi:10.1371/journal.pbio.0050121.
Lockless,S., Zhou, M., & MacKinnon, R.. Structural and Thermodynamic Properties of Selective Ion Binding in a K+ Channel. United States. doi:10.1371/journal.pbio.0050121.
Lockless,S., Zhou, M., and MacKinnon, R.. Mon . "Structural and Thermodynamic Properties of Selective Ion Binding in a K+ Channel". United States. doi:10.1371/journal.pbio.0050121.
@article{osti_960132,
title = {Structural and Thermodynamic Properties of Selective Ion Binding in a K+ Channel},
author = {Lockless,S. and Zhou, M. and MacKinnon, R.},
abstractNote = {Thermodynamic measurements of ion binding to the Streptomyces lividans K+ channel were carried out using isothermal titration calorimetry, whereas atomic structures of ion-bound and ion-free conformations of the channel were characterized by x-ray crystallography. Here we use these assays to show that the ion radius dependence of selectivity stems from the channel's recognition of ion size (i.e., volume) rather than charge density. Ion size recognition is a function of the channel's ability to adopt a very specific conductive structure with larger ions (K+, Rb+, Cs+, and Ba2+) bound and not with smaller ions (Na+, Mg2+, and Ca2+). The formation of the conductive structure involves selectivity filter atoms that are in direct contact with bound ions as well as protein atoms surrounding the selectivity filter up to a distance of 15 Angstroms from the ions. We conclude that ion selectivity in a K+ channel is a property of size-matched ion binding sites created by the protein structure.},
doi = {10.1371/journal.pbio.0050121},
journal = {PLoS Biology},
number = 5,
volume = 5,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 2007},
month = {Mon Jan 01 00:00:00 EST 2007}
}
  • The binding protein for the K{sup +}-channel toxin dendrotoxin I was purified from a detergent extract of rat brain membranes. The purification procedure utilized chromatography on DEAE-Trisacryl, affinity chromatography on a dendrotoxin-I-Aca 22 column, and wheat germ agglutinin-Affigel 10 with a final 3,800- to 4,600-fold enrichment and a recovery of 8-16%. The high affinity (K{sub d}, 40-100 pM) and specificity of the binding site are retained throughout the purification procedure. Analysis of the purified material on silver-stained NaDodSO{sub 4}/polyacrylamide gel revealed three bands of M{sub r} 76,000-80,000, 38,000 and 35,000. Interestingly, the binding site for {sup 125}I-labeled mast cell degranulatingmore » peptide, another putative K{sup +}-channel ligand from bee venom, which induces long-term potentiation in hippocampus, seems to reside on the same protein complex, as both binding sites copurify through the entire purification protocol.« less
  • Heulandite is a common rock-forming zeolite that exhibits wide solid solution of extra framework cations, presumably due to ready ion exchange with aqueous solutions. In order to provide a quantitative basis for interpreting and predicting the distribution of aqueous species between heulandite and aqueous solutions, ion exchange equilibrium between heulandite and aqueous solutions with respect to the binary cation pairs Ca{sup 2+} - K{sup +}, Ca{sup 2+} - Na{sup +}, K{sup +} - Na{sup +}, K{sup +} - Sr{sup 2+}, Na{sup +} - Sr{sup 2+}, and Ca{sup 2+} - Sr{sup 2+} was investigated. Homoionic Ca-, K-, and Na-heulandites prepared frommore » natural heulandite were equilibrated with 0.1 N Cl{sup -} solutions containing various proportions of the cations in a given binary pair at 55 and 85 C to define isotherms describing partitioning of the cations over a wide range of heulandite and solution composition with respect to the cations in each pair. In general, the experiments equilibrated rapidly, within 11-15 weeks at 55 C and 3-4 weeks at 85 C. The exception was the Ca{sup 2+} - Sr{sup 2+} binary exchange, which did not equilibrate even after 3 months at 55 C and 4 weeks at 85 C. Slow exchange of Sr{sup 2+} for Ca{sup 2+} also prohibited preparation of homoionic Sr-heulandite from the natural (Ca-rich) heulandite within 10 weeks in 2N SrCl{sub 2} solution at 90 C, although near homoionic Sr-heulandite was produced by exchange of K- and Na-heulandite. Experimentally determined isotherms were used to derive equilibrium constants for the ion exchange reactions and asymmetric Margules models describing the extent of non-ideality in extra framework solid solutions in heulandite. Regressed equilibrium constants for Ca{sup 2+}-Na{sup +}, Ca{sup 2+}-K{sup +}, and K{sup +}-Na{sup +} binary cation pairs at 55 C are internally consistent among each other (complying with the triangle rule), indicating good accuracy of these data. The maximum departure from internal Heulandite ion exchange Fridriksson and others consistency among the equilibrium constants for three binary pairs was 900 J per mole of charge equivalents (eq) for the 55 C experiments and 2300 J eq-1 for the 85 C experiments. The applicability of the present experimental results and thermodynamic models was assessed by calculating the composition of heulandite in Icelandic geothermal systems from known compositions using the regressed thermodynamic properties of Ca{sup 2+}-Na{sup +} exchange at 85 C. Calculations predict an average Ca mole fraction [defined as Ca/(Ca+Na)] in heulandite of 0.74, in excellent agreement with observed compositions of heulandite from geothermal and metamorphic systems in Iceland (0.75). Thermodynamic data for heulandite ion exchange derived in this study can be used to predict partitioning of Ca, K, Na, and Sr between heulandite and aqueous solutions in geologic systems. Because heulandite is the most effective sink for Sr in basaltic aquifers that have undergone zeolite facies metamorphism, the experimental results of this study will provide essential data for modeling Sr transport in aquifers in low-grade metabasalts.« less