X-ray structure of a ClC chloride channel at 3.0 Å reveals the molecular basis of anion selectivity

Dutzler, Raimund; Campbell, Ernest B.; Cadène, Martine; Chait, Brian T.; MacKinnon, Roderick

doi:10.1038/415287a

Cited by 1,507 publications

(1,852 citation statements)

References 46 publications

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“…Crystal structure and electrophysiological data from ClC channels clearly exclude permeation of ATP. The large organic anion ATP is much too big to pass through the ClC pore [62,63]. Likewise, high-resolution structural data from water and K + channels confirm that the narrow conductive pore is a general feature of ion channels [64,65].…”

Section: Suggested Mechanisms For Cellular Nucleotide Releasementioning

confidence: 89%

ATP release from non-excitable cells

Praetorius

Leipziger

2009

Purinergic Signalling

205

229

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All cells release nucleotides and are in one way or another involved in local autocrine and paracrine regulation of organ function via stimulation of purinergic receptors. Significant technical advances have been made in recent years to quantify more precisely resting and stimulated adenosine triphosphate (ATP) concentrations in close proximity to the plasma membrane. These technical advances are reviewed here. However, the mechanisms by which cells release ATP continue to be enigmatic. The current state of knowledge on different suggested mechanisms is also reviewed. Current evidence suggests that two separate regulated modes of ATP release co-exist in nonexcitable cells: (1) a conductive pore which in several systems has been found to be the channel pannexin 1 and (2) vesicular release. Modes of stimulation of ATP release are reviewed and indicate that both subtle mechanical stimulation and agonist-triggered release play pivotal roles. The mechano-sensor for ATP release is not yet defined.

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Section: Suggested Mechanisms For Cellular Nucleotide Releasementioning

confidence: 89%

ATP release from non-excitable cells

Praetorius

Leipziger

2009

Purinergic Signalling

205

229

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“…Nonetheless, the contributions to g dehydr do not solely originate from the alkyl tail, depending on the chemical nature of the headgroup. Finally, the hydration free energy term g hydr in eqn (22) accounts for free energy changes experienced by water molecules that remain in a hydrating contact with the detergent upon micelle formation.…”

Section: Cs-mt Approach To Cmentioning

confidence: 99%

“…Rather, the focus is on the effective interactions between detergent micelles and protein-detergent complexes (PDCs) induced by PEG. The interest in these interactions originates from the seminal work of George and Wilson 30 who used the second osmotic virial coefficient (B 22 ) as a measure of intermolecular potential to define crystallization conditions for soluble proteins. They found the B 22 values under crystallization conditions to fall into a fairly narrow range below zero, which they referred to as the ''crystallization slot'' and which indicates small attractive interactions between protein molecules.…”

Section: Introductionmentioning

confidence: 99%

The influence of poly(ethylene glycol) on the micelle formation of alkyl maltosides used in membrane protein crystallization

Müh

DiFiore

Zouni

2015

Phys. Chem. Chem. Phys.

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With the aim of better understanding the phase behavior of alkyl maltosides (n-alkyl-b-D-maltosides, C n G 2 ) under the conditions of membrane protein crystallization, we studied the influence of poly(ethylene glycol) (PEG) 2000, a commonly used precipitating agent, on the critical micelle concentration (CMC) of the alkyl maltosides by systematic variation of the number n of carbon atoms in the alkyl chain (n = 10, 11, and 12) and the concentration of PEG2000 (w) in a buffer suitable for the crystallization of cyanobacterial photosystem II. CMC measurements were based on established fluorescence techniques using pyrene and 8-anilinonaphthalene-1-sulfonate (ANS). We found an increase of the CMC with increasing PEG concentration according to ln(CMC/CMC 0 ) = k P w, where CMC 0 is the CMC in the absence of PEG and k P is a constant that we termed the ''polymer constant''. In parallel, we measured the influence of PEG2000 on the surface tension of detergent-free buffer solutions. At PEG concentrations w 4 1% w/v, the surface pressure p s (w) = g(0) À g(w) was found to depend linearly on the PEG concentration according to p s (w) = kw + p s (0), where g (0) is the surface tension in the absence of PEG. Based on a molecular thermodynamic modeling, CMC shifts and surface pressure due to PEG are related, and it is shown that k P = kc(n) + Z, where c(n) is a detergent-specific constant depending inter alia on the alkyl chain length n and Z is a correction for molarity. Thus, knowledge of the surface pressure in the absence of a detergent allows for the prediction of the CMC shift. The PEG effect on the CMC is discussed concerning its molecular origin and its implications for membrane protein solubilization and crystallization.

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“…Each subunit consists of 18 a-helical domains designated A-R (12)(13)(14). Eukaryotic and some prokaryotic CLC proteins also have large cytoplasmic C-termini containing two cystathionine-b-synthase (CBS) domains, termed CBS1 and CBS2, that dimerize to form a Bateman domain (14)(15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Role of CBS and Bateman Domains in Phosphorylation-Dependent Regulation of a CLC Anion Channel

Yamada

Krzeminski

Bozóky

et al. 2016

Biophysical Journal

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Eukaryotic CLC anion channels and transporters are homodimeric proteins composed of multiple a-helical membrane domains and large cytoplasmic C-termini containing two cystathionine-b-synthase domains (CBS1 and CBS2) that dimerize to form a Bateman domain. The Bateman domains of adjacent CLC subunits interact to form a Bateman domain dimer. The functions of CLC CBS and Bateman domains are poorly understood. We utilized the Caenorhabditis elegans CLC-1/2/Ka/ Kb anion channel homolog CLH-3b to characterize the regulatory roles of CLC cytoplasmic domains. CLH-3b activity is reduced by phosphorylation or deletion of a 14-amino-acid activation domain (AD) located on the linker connecting CBS1 and CBS2. We demonstrate here that phosphorylation-dependent reductions in channel activity require an intact Bateman domain dimer and concomitant phosphorylation or deletion of both ADs. Regulation of a CLH-3b AD deletion mutant is reconstituted by intracellular perfusion with recombinant 14-amino-acid AD peptides. The sulfhydryl reactive reagent 2-(trimethylammonium)ethyl methanethiosulfonate bromide (MTSET) alters in a phosphorylation-dependent manner the activity of channels containing single cysteine residues that are engineered into the short intracellular loop connecting membrane a-helices H and I (H-I loop), the AD, CBS1, and CBS2. In contrast, MTSET has no effect on channels in which cysteine residues are engineered into intracellular regions that are dispensable for regulation. These studies together with our previous work suggest that binding and unbinding of the AD to the Bateman domain dimer induces conformational changes that are transduced to channel membrane domains via the H-I loop. Our findings provide new, to our knowledge, insights into the roles of CLC Bateman domains and the structurefunction relationships that govern the regulation of CLC protein activity by diverse ligands and signaling pathways.

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X-ray structure of a ClC chloride channel at 3.0 Å reveals the molecular basis of anion selectivity

Cited by 1,507 publications

References 46 publications

ATP release from non-excitable cells

ATP release from non-excitable cells

The influence of poly(ethylene glycol) on the micelle formation of alkyl maltosides used in membrane protein crystallization

Role of CBS and Bateman Domains in Phosphorylation-Dependent Regulation of a CLC Anion Channel

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