Structural Basis for Differences in Dynamics Induced by Leu Versus Ile Residues in the CD Loop of Kir Channels

Lü, Shouqin; An, Hailong; Zhang, Hailin; Long, Mian

doi:10.1007/s12035-015-9466-x

Cited by 5 publications

(7 citation statements)

References 34 publications

(76 reference statements)

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“…The most interesting observed pocket is a conservative site, consisting in physical gates formed by cytoplasmic (G loop) region ( Hibino et al, 2010 ), necessary for the regulation of the inward potassium current that obviously depends on the state of the gate ( Lü et al, 2016 ). The G loop has a crucial role with their amino acid composition in the regulation of K ATP gating kinetics, different previous works shown how mutations of the amino acid of this region are responsible of protein inactivation ( Shimomura et al, 2009 ; Li et al, 2016 ; Nishida et al, 2007 ; Pegan et al, 2006 ; Hattersley & Ashcroft, 2005 ; Proks et al, 2005 ).…”

Section: Resultsmentioning

confidence: 99%

“…Thus, the inherent dynamics and structural stability of TM and CTDs were investigated. The initially applied harmonic restraints on the protein backbone were gradually released during the course of the second 1 ns simulation followed by additional 50 ns MD simulations without any restraints ( Ismail & Jusoh, 2016 ; Haider et al, 2007 ; Lü et al, 2016 ), specifying that the topology of the channel was maintained under the condition of no restraints ( Figs. 5 and 6 ).…”

Section: Resultsmentioning

confidence: 99%

“…3 ), in order to propose a potential consensus binding site of Kir6.1 and a flavonoid mechanism of action inside the binding pocket. Through bioinformatics approaches and previous works ( Hibino et al, 2010 ), we identified the G loop as potential binding site for ligands ( Lü et al, 2016 ; Shimomura et al, 2009 ; Li et al, 2016 ; Nishida et al, 2007 ; Pegan et al, 2006 ; Hattersley & Ashcroft, 2005 ; Proks et al, 2005 ). Docking simulation and different MD simulation protocols were accomplished for explaining how the flavonoids influenced the gating process of the channel.…”

Section: Discussionmentioning

confidence: 99%

“…The TMD and CTD are linked by about 20 amino acids and four supporting proteins, the sulfonylurea receptors, which envelop the four subunits of K ATP forming a hetero-octameric complex ( Sepúlveda et al, 2015 ). The Kir channel open-closed mechanism depends on different conformational changes which regulate its state ( Li et al, 2016 ; Lü et al, 2016 ). Small molecules regulate functions of Kir channels for instance: H + , Mg 2+ , Na + ; polyamines, phosphorylation and membrane-bound phospholipids and proteins ( Xie et al, 2007 ; Hibino et al, 2010 ; Li et al, 2016 ; Fowler et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

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From in silico to in vitro: a trip to reveal flavonoid binding on the Rattus norvegicus Kir6.1 ATP-sensitive inward rectifier potassium channel

Trezza

Cicaloni

Porciatti

et al. 2018

PeerJ

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BackgroundATP-sensitive inward rectifier potassium channels (Kir), are a potassium channel family involved in many physiological processes. KATP dysfunctions are observed in several diseases such as hypoglycaemia, hyperinsulinemia, Prinzmetal angina–like symptoms, cardiovascular diseases.MethodsA broader view of the KATP mechanism is needed in order to operate on their regulation, and in this work we clarify the structure of the Rattus norvegicus ATP-sensitive inward rectifier potassium channel 8 (Kir6.1), which has been obtained through a homology modelling procedure. Due to the medical use of flavonoids, a considerable increase in studies on their influence on human health has recently been observed, therefore our aim is to study, through computational methods, the three-dimensional (3D) conformation together with mechanism of action of Kir6.1 with three flavonoids.ResultsComputational analysis by performing molecular dynamics (MD) and docking simulation on rat 3D modelled structure have been completed, in its closed and open conformation state and in complex with Quercetin, 5-Hydroxyflavone and Rutin flavonoids. Our study showed that only Quercetin and 5-Hydroxyflavone were responsible for a significant down-regulation of the Kir6.1 activity, stabilising it in a closed conformation. This hypothesis was supported by in vitro experiments demonstrating that Quercetin and 5-Hydroxyflavone were capable to inhibit KATP currents of rat tail main artery myocytes recorded by the patch-clamp technique.ConclusionCombined methodological approaches, such as molecular modelling, docking and MD simulations of Kir6.1 channel, used to elucidate flavonoids intrinsic mechanism of action, are introduced, revealing a new potential druggable protein site.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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From in silico to in vitro: a trip to reveal flavonoid binding on the Rattus norvegicus Kir6.1 ATP-sensitive inward rectifier potassium channel

Trezza

Cicaloni

Porciatti

et al. 2018

PeerJ

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“…Positively charged residues K182, K185 and K187 on Kir2.1 inner TMD and the tether helix structure (equivalent to K183, R186, and K188 in Kir2.2), are candidates for such interaction. These residues directly interact with 5′ phosphate in the inositol ring of PIP 2 in a Kir2.2-PIP 2 structural model (Hansen et al, 2011;Lu et al, 2016). We performed a limited structural analysis of Kir2.1 valine 77 substitution to glutamic acid, based on Kir2.2-PIP 2 crystal structure (PDB ID: 3SPI) where the corresponding residue is valine 75.…”

Section: Structural Analysis Of the Mutated Kir21mentioning

confidence: 99%

Andersen–Tawil Syndrome Is Associated With Impaired PIP2 Regulation of the Potassium Channel Kir2.1

et al. 2020

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Andersen-Tawil syndrome (ATS) type-1 is associated with loss-of-function mutations in KCNJ2 gene. KCNJ2 encodes the tetrameric inward-rectifier potassium channel Kir2.1, important to the resting phase of the cardiac action potential. Kir-channels' activity requires interaction with the agonist phosphatidylinositol-4,5-bisphosphate (PIP 2). Two mutations were identified in ATS patients, V77E in the cytosolic N-terminal "slide helix" and M307V in the C-terminal cytoplasmic gate structure "G-loop." Current recordings in Kir2.1-expressing HEK cells showed that each of the two mutations caused Kir2.1 lossof-function. Biotinylation and immunostaining showed that protein expression and trafficking of Kir2.1 to the plasma membrane were not affected by the mutations. To test the functional effect of the mutants in a heterozygote set, Kir2.1 dimers were prepared. Each dimer was composed of two Kir2.1 subunits joined with a flexible linker (i.e. WT-WT, WT dimer; WT-V77E and WT-M307V, mutant dimer). A tetrameric assembly of Kir2.1 is expected to include two dimers. The protein expression and the current density of WT dimer were equally reduced to~25% of the WT monomer. Measurements from HEK cells and Xenopus oocytes showed that the expression of either WT-V77E or WT-M307V yielded currents of only about 20% compared to the WT dimer, supporting a dominant-negative effect of the mutants. Kir2.1 sensitivity to PIP 2 was examined by activating the PIP 2 specific voltage-sensitive phosphatase (VSP) that induced PIP 2 depletion during current recordings, in HEK cells and Xenopus oocytes. PIP 2 depletion induced a stronger and faster decay in Kir2.1 mutant dimers current compared to the WT dimer. BGP-15, a drug that has been demonstrated to have an anti-arrhythmic effect in mice, stabilized the Kir2.1 current amplitude following VSP-induced PIP 2 depletion in cells expressing WT or mutant dimers. This study underlines the implication of mutations in cytoplasmic regions of Kir2.1. A newly developed calibrated VSP activation protocol enabled a quantitative assessment of changes in PIP 2 regulation caused by the mutations. The results suggest an impaired function and a dominant-negative effect of the Kir2.

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Inhibition of inwardly rectifying Kir2.x channels by the novel anti-cancer agent gambogic acid depends on both pore block and PIP2 interference

Scherer

Schworm

Seyler

et al. 2017

Naunyn-Schmiedeberg's Arch Pharmacol

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The caged xanthone gambogic acid (GA) is a novel anti-cancer agent which exhibits anti-proliferative, anti-inflammatory and cytotoxic effects in many types of cancer tissues. In a recent phase IIa study, GA exhibits a favourable safety profile. However, limited data are available concerning its interaction with cardiac ion channels. Heteromeric assembly of Kir2.x channels underlies the cardiac inwardly rectifying I current which is responsible for the stabilization of the diastolic resting membrane potential. Inhibition of the cardiac I current may lead to ventricular arrhythmia due to delayed afterdepolarizations. Compared to Kv2.1, hERG and Kir1.1, a slow, delayed inhibition of Kir2.1 channels by GA in a mammalian cell line was reported before but no data exist in literature concerning action of GA on homomeric Kir2.2 and Kir2.3 and heteromeric Kir2.x channels. Therefore, the aim of this study was to provide comparative data on the effect of GA on homomeric and heteromeric Kir2.x channels. Homomeric and heteromeric Kir2.x channels were heterologously expressed in Xenopus oocytes, and the two-microelectrode voltage-clamp technique was used to record Kir2.x currents. To investigate the mechanism of the channel inhibition by GA, alanine-mutated Kir2.x channels with modifications in the channels pore region or at phosphatidylinositol 4,5-bisphosphate (PIP)-binding sites were employed. GA caused a slow inhibition of homomeric and heteromeric Kir2.x channels at low micromolar concentrations (with IC Kir2.1/2.2 < Kir2.2 < Kir2.2/2.3 < Kir2.3 < Kir2.1 < Kir2.1/2.3). The effect did not reach saturation within 60 min and was not reversible upon washout for 30 min. The inhibition showed no strong voltage dependence. We provide evidence for a combination of direct channel pore blockade and a PIP-dependent mechanism as a molecular basis for the observed effect. We conclude that Kir2.x channel inhibition by GA may be relevant in patients with pre-existing cardiac disorders such as chronic heart failure or certain rhythm disorders and recommend a close cardiac monitoring for those patients when treated with GA.

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Structural Basis for Differences in Dynamics Induced by Leu Versus Ile Residues in the CD Loop of Kir Channels

Cited by 5 publications

References 34 publications

From in silico to in vitro: a trip to reveal flavonoid binding on the Rattus norvegicus Kir6.1 ATP-sensitive inward rectifier potassium channel

From in silico to in vitro: a trip to reveal flavonoid binding on the Rattus norvegicus Kir6.1 ATP-sensitive inward rectifier potassium channel

Andersen–Tawil Syndrome Is Associated With Impaired PIP2 Regulation of the Potassium Channel Kir2.1

Inhibition of inwardly rectifying Kir2.x channels by the novel anti-cancer agent gambogic acid depends on both pore block and PIP2 interference

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