Open conformation of hERG channel turrets revealed by a specific scorpion toxin BmKKx2
© Hu et al.; licensee BioMed Central Ltd. 2014
Received: 3 January 2014
Accepted: 11 February 2014
Published: 11 April 2014
The human ether-a-go-go-related gene potassium channel (hERG) has an unusual long turret, whose role in recognizing scorpion toxins remains controversial. Here, BmKKx2, the first specific blocker of hERG channel derived from scorpion Mesobuthus martensii, was identified and the turret role of hERG channel was re-investigated using BmKKx2 as a molecular probe.
BmKKx2 was found to block hERG channel with an IC50 of 6.7 ± 1.7 nM and share similar functional surface with the known hERG channel inhibitor BeKm-1. The alanine-scanning mutagenesis data indicate that different residue substitutions on hERG channel by alanine decreased the affinities of toxin BmKKx2 by about 10-fold compared with that of wild-type hERG channel, which reveals that channel turrets play a secondary role in toxin binding. Different from channel turret, the pore region of hERG channel was found to exert the conserved and essential function for toxin binding because the mutant hERG-S631A channel remarkably decreased toxin BmKKx2 affinity by about 104-fold.
Our results not only revealed that channel turrets of hERG channel formed an open conformation in scorpion toxin binding, but also enriched the diversity of structure-function relationships among the different potassium channel turrets.
Potassium channels mediate K+ efflux and play various pharmacological functions. Structurally, the potassium channels present various topologies, such as inwardly rectifying two-transmembrane potassium channels (Kir), calcium-activated potassium channels with six or seven transmembrane segments (KCa) and voltage-gated potassium channels with six transmembrane segments (Kv). Due to the channel structure complexity and technique challenge, there are a few crystal structures of eukaryotic potassium channels, such as Kv1.2, Kir2.2 and Kir3.2[3–5]. Comparison of these crystal structures reveals essential conformation differences in the extracellular pore entryway that includes turret and filter regions. In recent years, other structural features of channel turrets were also revealed by the specific scorpion toxins as the molecular probes. For example, the 10-residue turrets of Kv1.2 channel is induced to form an open state conformation and not affect toxin binding activity when it is bound by scorpion toxin maurotoxin. Different from the open state conformation of Kv1.2 channel turret, the 17-residue turrets of small conductance KCa3 (SKCa3) channel form a “peptide screener” compact conformation which selectively controls scorpion toxin binding. More interestingly, the 19-residue turret of large conductance KCa (BKCa) channel switches from an open state conformation for scorpion toxin charybdotoxin (ChTX) binding to a compact “helmet” conformation for toxin ChTX insensitivity when the channel turret interacts with its auxiliary β4 subunits. Besides above progress of structure and function relationships of different potassium channel turrets, characterization of more potassium channel extracellular pore entryways remains a challenge nowadays due to the difficulties in determining the crystal structures of additional potassium channels.
Among all the superfamily members of potassium channels, hERG (human ether-a-go-go-related gene) potassium channel has an unusual longer turret containing 40 amino acid residues. Using the scorpion toxin BeKm-1 as a molecular probe, different conformational states in the turret of hERG channel have been reported. Through the computational simulation technique, our group ever predicted that hERG channel vestibule presented an open conformation mainly in a decentralized ‘petunia’ shape and turrets were far from the bound toxin BeKm-1, while another model without a ‘petunia’ shape was also proposed by other group, in which channel turrets directly interacted with the bound BeKm-1. Thus, these controversial structural-functional features still need to be investigated using scorpion toxins as a molecular probe. In this work, we revisit the conformational state of hERG channel turrets by using a specific scorpion toxin BmKKx2, a novel toxin from Scorpion Mesobuthus martensii with similar structural and functional features as BeKm-1. Through alanine-scanning mutagenesis and pharmacological experiments, hERG channel pore region instead of turrets was mainly responsible for toxin BmKKx2 binding, supporting our previous inference that the hERG channel turret adopts an open conformation. Taken together, these findings provide new insights into the role of channel turret in toxin recognition and might help to better understand the structure-function relationship of hERG channel.
Expression and purification of BmKKx2
The GST-BmKKx2, a fusion protein of 30 kDa size, was purified and split into two products, GST of 26 kDa and BmKKx2 of 4.1 kDa (Figure 1B). The mixture was further separated by HPLC, and two product peaks were obtained (Figure 1C). The component eluting at about 18 min corresponding to BmKKx2 was collected manually and lyophilized. By the matrix-assisted-laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), the molecular weight of recombinant BmKKx2 was 3999.6 (Figure 1D). Considered the loss of 6 Da by three pairs of the conserved disulfide bridges, the determined molecular weight was in good agreement with the theoretical molecular weight of 4005.6 Da calculated by the ExPASy Protemics Server (http://us.expasy.org/tools/protparam.html).
BmKKx2 appears to be a selective hERG channel blocker
The binding affinities of wild-type BmKKx2 and its mutants towards hERG channel
26.7 ± 1.7
238.3 ± 11.2
131.0 ± 35.2
376.6 ± 62.0
409.6 ± 56.9
863.4 ± 73.4
123.2 ± 56.3
262.7 ± 25.9
24.1 ± 1.9
Less effect of channel turret on BmKKx2 binding
The binding affinities of toxin BmKKx2 towards wild-type and mutant hERG channel
6.7 ± 1.7
76.5 ± 32.6
87.5 ± 16.8
24.3 ± 8.4
59.7 ± 31.7
14.5 ± 4.3
19.6 ± 1.7
89.5 ± 39.8
65.2 ± 19.1
60.8 ± 23.4
118.2 ± 26.7
15.2 ± 4.8
12.1 ± 3.5
116.1 ± 29.2
6.7 ± 5.1
42.6 ± 17.1
698.0 ± 64.4
Secondly, we continued to investigate the effect of the controversial helix region in channel turrets on toxin BeKm-1 binding[10, 11]. Except no expression of hERG-W585A and lower expression level of hERG-L589A, other eight mutant channels (hERG-L586A, hERG-H587A, hERG-N588A, hERG-D591A, hERG-Q592A, hERG-K595A, hERG-P596A and hERG-Y597A) exhibited the similar properties to that of wild-type hERG channels, and their effects on toxin BmKKx2 binding were determined (Figure 5C-J). The corresponding IC50 values of toxin BmKKx2 were 24.3, 59.7, 14.5, 19.6, 89.5, 65.2, 60.8 and 118.2 nM for hERG-L586A, hERG-H587A, hERG-N588A, hERG-D591A, hERG-Q592A, hERG-K595A, hERG-P596A and hERG-Y597A, respectively (Figure 6A-C and Table 2). Overall, there were drops of no more than 20-fold toxin sensitivity towards eight mutant hERG channels, which indicated that the helix region in channel turrets did not play an important role in scorpion toxin recognition.
Finally, the influence of the HP linker in channel turret in BmKKx2 recognition was also studied. Six mutant channels were constructed, and mutant hERG-D609A channel could not be investigated due to the negligibly small currents. Other five mutant channels (hERG-N598A, hERG-S600A, hERG-K608A, hERG-K610A and hERG-T613A) were able to mediate potassium currents with comparable amplitude with wild-type hERG channel, and also were effectively blocked by 100 nM toxin BmKKx2 (Figure 5K-O). The IC50 values of toxin BmKKx2 for hERG-N598A, hERG-S600A, hERG-K608A, hERG-K610A and hERG-T613A were 15.2, 12.1, 116.1, 6.7 and 42.6 nM, respectively (Figure 6C and D). Similar to the previous two S5H and helix regions of the channel turret (Table 2), these experimental data of HP linker residues indicated that it also played a secondary role in the toxin BmKKx2 binding.
Critical role of channel pore region in BmKKx2 binding
The interaction between animal toxin blockers and potassium channels indicated that the channel pore region played a critical role in toxin recognition[19, 20]. In this work, two residues Asn629 and Ser631 near the channel selectivity filter were selected and substituted by alanine residue, respectively. Since the lower expression of the mutant channel hERG-N629A, only mutant hERG-S631A channel was investigated for toxin BmKKx2 recognition. As shown in Figure 5P, mutant hERG-S631A channel currents could be inhibited by 100 nM toxin BmKKx2, and its IC50 of toxin BmKKx2 was 698 ± 64.4 nM, making hERG-S631A approximately 104-fold less sensitive to BmKKx2 than wild-type hERG channel (Figure 6D). The remarkable effect of channel Ser631 on toxin binding confirmed that the channel pore region was greatly involved in toxin binding.
With about 100 members, the potassium channels present the diverse structure-function relationships. The extracellular pore entryway including turrets and filter regions is the critical domain responsible for the inactivation, animal toxin recognition and interactions between channel subunits. Due to the difficulties in solving the crystal structures of potassium channels, the scorpion toxin blockers are useful molecular probes to characterize the structure-function relationships of the different channel extracellular pore entryways[6–8, 20].
In this work, we revisited the hERG channel turret with an unusual longer turret containing 40 amino acid residues (Figure 4), whose role was ever intensively investigated in the scorpion toxin binding[10, 11, 17]. However, there were two different inferences on the orientation of hERG channel turrets: one is the helix domain of hERG channel turrets responsible for scorpion toxin binding through the cysteine-scanning mutagenesis and computational modeling, the other is the turret helix domain far from the bound scorpion toxin through the computational modeling. Unexpectedly, the cysteine substitutions of hERG channel turrets at different positions were found to have high-impact, intermediate-impact or low-impact on channel functions and affect scorpion toxin binding due to the potential formation of the disulfide bonds[11, 22]. In order to avoid the effect of disulfide bonds on scorpion toxin binding, the alanine-scanning mutagenesis was used to re-investigate the role of hERG channel turret, especially the controversial turret helix domain, in the scorpion toxin binding.
Conserved functional surface of new toxin BmKKx2
Open conformation of hERG channel turrets in toxin BmKKx2 recognition
As the role of channel turrets in scorpion toxin binding, the controversial issue was whether the helix domain interacted with the bound toxin (Figure 4)[10, 11] although this helix domain actually did not exist due to locating above the extracellular pore entryway in the aqueous solution instead of the hydrophobic environment. In previous cysteine-scanning experiments, total 29 mutant channels were investigated while their cysteine substitutions had intermediate-impact or low-impact on the channel function. It was found that the substitution of Gln592 by the cysteine in the helix domain significantly decreased the affinity of toxin BeKm-1 by about 54-fold, and the remaining 28 turret residue substitutions including three S5H, helix and HP regions (Figure 4) decreased toxin BeKm-1 affinities by less than 10-fold when compared with the that of wild-type hERG channel. These results supported the inference that helix domain of channel turrets would interact with toxin BeKm-1. In our work, 15 channel turret residues were successfully assessed through the alanine-scanning mutagenesis (Figures 5 and6, and Table 2). Different from the important effect of channel turret Gln592 on toxin BeKm-1 binding, there was only about 13-fold drop in toxin BmKKx2 binding affinity when Gln592 residue was replaced by alanine (Figures 5G,6B and Table 2). Since the alanine has shorter sidechain than that of cysteine, and the substitution of channel turret Gln592 by cysteine was found to have intermediate-impact on hERG channel function shown by the differential effects of mutant channel function in the presence or absence of reducing agent DTT. In combination with the less effect of mutant hERG-Q592A on toxin BmKKx2 affinity (Figures 5G,6B and Table 2), channel turret Gln592 did not play a critical role in scorpion toxin BeKm-1 or BmKKx2 binding since toxin BeKm-1 or BmKKx2 had the similar structures and common functional residues (Figure 7A and B).
Further comparison of the pharmacological data of toxin BmKKx2 and BeKm-1 towards mutant hERG channels showed a common feature that three turret regions including S5H, helix and HP played an unimportant role in toxin binding (Figures 4,5, and6 and Table 2). In our previous work, the hERG channel turrets were predicted to possibly affect toxin BeKm-1 binding process because of its significant flexibility although they were likely far from the bound toxin BeKm-1 during the molecular dynamic simulations. This inference was supported by about 10-fold drops of toxin BmKKx2 affinities towards mutant hERG-R582A, hERG-I583A, hERG-H587A, hERG-Q592A, hERG-Y597A, hERG-K608A (Figures 5,6 and Table 2).
In summary, the comprehensive analysis of the pharmacological data of toxin BmKKx2 and BeKm-1 towards mutant hERG channels showed that channel turrets formed an open conformation in scorpion toxin binding (Figures 5,6 and Table 2). This open conformation was further shown in the toxin BmKKx2-hERG channel complex through the structural modeling using our previous toxin BeKm-1-hERG channel complex as the template (Figure 7C). Such conformational feature of hERG channel turrets was also observed in the turrets of Kv1.2 channel bound by scorpion toxin maurotoxin and the turrets of BKCa channel bound by scorpion toxin ChTX.
Conserved function of hERG channel pore region in toxin BmKKx2 recognition
The pore regions of potassium channels play an important role during its interactions with the channel-blocking animal toxins[19, 20]. In this work, the channel Ser631 was found essential for toxin BmKKx2 binding shown by about 104-fold drop of BmKKx2 affinity compared with that of wild-type hERG channel (Figures 5P,6D and Table 2). As shown in Figure 4, the corresponding residues in the position of Ser631 near hERG channel selectivity filter were critical for toxin activities among different potassium channels[6, 19, 23], which also rationalized the conserved and critical function of Ser631 in the hERG channel pore region. In the previous cysteine-scanning experiments, the importance of Ser631 residue in hERG channel was less significant likely since its substitution by cysteine was found to have intermediate-impact on hERG channel function shown by the differential effects of mutant channel function in the presence or absence of reducing agent DTT. As shown in Figure 7D, the polar and nonpolar interactions between channel Ser631 and toxin residues further illustrated the essential function of channel Ser631 for toxin binding in the modeled hERG channel-toxin BmKKx2 complex structure.
In this work, we revisited the hERG potassium channel which has an unusual longer turret with the considerable conformational flexibility. With the help of a new specific scorpion toxin BmKKx2, the first specific blocker for hERG channels from scorpion Mesobuthus martensii, the turret role of hERG channel was re-investigated through the alanine-scanning mutagenesis. All pharmacological data indicated that different residue substitutions by alanine decreased the affinities of toxin BmKKx2 by about 10-fold compared with that of wild-type hERG channel, which revealed that channel turret played a secondary role in toxin binding. The further comprehensive analysis of the pharmacological data of scorpion toxins towards mutant hERG channels from the previous cysteine-scanning and present alanine-scanning mutagenesis showed that channel turrets formed an open conformation in scorpion toxin binding. Together, these findings were helpful to understand the unique role of hERG channel turrets in toxin recognition and enrich the diversity of turret structure-function relationships among the different potassium channels.
Clone, expression and purification of BmKKx2 peptide
The cDNA library of Mesobuthus martensii was constructed by our group as described previously. cDNAs were cloned into pSPORT1 plasmids and transformed into E. coli DH5α cells (obtained from China Center for Type Culture Collection, CCTCC). Randomly chosen cDNA clones were sequenced to obtain a reliable representation of the toxin content in the venom gland.
After acquiring the full-length cDNA of BmKKx2, expression plasmid pGEX-6p-1-BmKKx2 was constructed using overlap PCR method. The primers used were: forward primer 1: 5’-CCTACAGATATTAAATGCAGTGCATCTTATCAATGT- TTCCCTGTTTG-3’; reverse primer 1: 5’-AACGTCCATTAGTCTTTCCGAAGCG-GCTTTTACAAACAGGGAAACATTG-3’; forward primer 2: 5’-GTGAATTCGAT-GACGATGACAAG CGTCCTACAGATATTAAATG-3’; reverse primer 2: 5’-TAGCTCGAGCTAGAAACAGTCGCATAAACCATTCACGCAACGTCCATTAG-3’. The restriction enzyme sites are underlined, and an enterokinase cleavage site is in bold. The PCR products were inserted into expression vector pGEX-6p-1 and sequenced with universal pGEX primers. E. coli Rosetta (DE3) cells were used to express BmKKx2 and its mutants according to previous techniques of our group[7, 10, 15, 21].
Alanine-scanning mutagenesis of hERG channel
The wild-type hERG-pEGFP-N2 vector was used in this work. Twenty-two mutants were chosen for study base on the structure analysis of hERG channel and our previous model. All the channel mutants were constructed with the QuickChange site-directed mutagenesis kit (Stratagene), as described by the manufacture. The mutated expression plasmids were confirmed by sequencing after construction (Takara).
Cell culture and transfection
Human embryonic kidney (HEK293) cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM, Gibco) with 10% Fetal Calf Serum (Invitrogen Life Technologies, Carlesbad, CA), supplemented with 100 units/mL ampicillin and 100 μg/mL streptomycin in a humidified 5% CO2 incubator at 37°C.
Cells were transfected with 6 μg plasmid of wild-type or mutant hERG using the FuGene Transfection Method (Roche Diagnostics, Switzerland). Currents were recorded 24–48 hours after transfection in EGFP fluorescent cells at room temperature using the whole-cell patch-clamp mode with an EPC10 Amplifier (HEKA, Germany).
Electrophysiology and data analysis
Cells expressing Kv channels were incubated in the external solution consisted of (in mM): 137 NaCl, 4 KCl, 1 MgCl2, 1.8 CaCl2, 10 D-Glucose and 10 HEPES (pH7.4 with NaOH); and the internal solution contained (in mM): 130 KCl, 1 MgCl2, 5 MgATP, 5 EGTA, 10 HEPES (pH 7.2 with KOH). For Kv1 channels and Kv4.2, currents were elicited by 200-ms or 500-ms depolarizing pulses from a holding potential of -80 to 50 mV, individually. While cells expressing hERG channel were depolarized from a holding potential of -80 mV to +40 mV for 500 ms then hyperpolarized to -120 mV for 1 s and current amplitudes were measured from the peak inward current at -120 mV.
For the recording of SKCa channels, the bath solution contained 130 mM sodium aspartate, 30 mM K+-aspartate, 2 mM CaCl2, 1 mM MgCl2, and 10 mM HEPES (pH 7.4 with NaOH). The pipette solution contained 145 mM K+-aspartate, 8.7 mM CaCl2, 2 mM MgCl2, 10 mM EGTA, and 10 mM HEPES (pH 7.2 with KOH) to achieve an intracellular free Ca2+ concentration of 1 μM. The membrane potentials were clamped to -120 mV for 50 ms (which was used for the current measurements), followed by a 400 ms voltage ramp from -120 to +60 mV and were kept for 5 s between ramps at -40 mV.
where IC50 is the half-maximum inhibition concentration, I is the peak tail current, [T] represents the concentration of toxins. Results are shown as mean ± S.E., and n is the number of experiments.
This work was supported by grants from the National Basic Research Program of China (No. 2010CB529800), National High Technology Research and Development Program of China (No. 2012AA020304), the National Natural Sciences Foundation of China (No.31170789), the New Century Excellent Talents in Wuhan University by the Ministry of Education of China (No. NCET-10-0651) and Wuhan City Science and Technology Foundation of China (No. 2013070204020046).
- Ashcroft FM: From molecule to malady. Nature. 2006, 440: 440-447. 10.1038/nature04707View ArticlePubMedGoogle Scholar
- Shieh CC, Coghlan M, Sullivan JP, Gopalakrishnan M: Potassium channels: molecular defects, diseases, and therapeutic opportunities. Pharmacol Rev. 2000, 52: 557-594.PubMedGoogle Scholar
- Long SB, Campbell EB, Mackinnon R: Crystal structure of a mammalian voltage-dependent Shaker family K+ channel. Science. 2005, 309: 897-903. 10.1126/science.1116269View ArticlePubMedGoogle Scholar
- Tao X, Avalos JL, Chen J, MacKinnon R: Crystal structure of the eukaryotic strong inward-rectifier K+ channel Kir2.2 at 3.1 Å resolution. Science. 2009, 326: 1668-1674. 10.1126/science.1180310PubMed CentralView ArticlePubMedGoogle Scholar
- Whorton MR, MacKinnon R: X-ray structure of the mammalian GIRK2-betagamma G-protein complex. Nature. 2013, 498: 190-197. 10.1038/nature12241PubMed CentralView ArticlePubMedGoogle Scholar
- Yi H, Qiu S, Cao ZJ, Wu YL, Li WX: Molecular basis of inhibitory peptide maurotoxin recognizing Kv1.2 channel explored by ZDOCK and molecular dynamic simulations. Proteins. 2008, 70: 844-854.View ArticlePubMedGoogle Scholar
- Feng J, Hu YT, Yi H, Yin SJ, Han S, Hu J, Chen ZY, Yang WS, Cao ZJ, De Waard M, Sabatier JM, Li WX, Wu YL: Two conserved arginine residues from the SK3 potassium channel outer vestibule control selectivity of recognition by scorpion toxins. J Biol Chem. 2013, 288: 12544-12553. 10.1074/jbc.M112.433888PubMed CentralView ArticlePubMedGoogle Scholar
- Gan GL, Yi H, Chen MR, Sun L, Li WX, Wu YL, Ding JP: Structural basis for toxin resistance of beta4-associated calcium-activated potassium (BK) channels. J Biol Chem. 2008, 283: 24177-24184. 10.1074/jbc.M800179200PubMed CentralView ArticlePubMedGoogle Scholar
- Torres AM, Bansal PS, Sunde M, Clarke CE, Bursill JA, Smith DJ, Bauskin A, Breit SN, Campbell TJ, Alewood PF, Kuchel PW, Vandenberg JI: Structure of the HERG K+ channel S5P extracellular linker: role of an amphipathic alpha-helix in C-type inactivation. J Biol Chem. 2003, 278: 42136-42148. 10.1074/jbc.M212824200View ArticlePubMedGoogle Scholar
- Yi H, Cao ZJ, Yin SJ, Dai C, Wu YL, Li WX: Interaction simulation of hERG K+ channel with its specific BeKm-1 peptide: insights into the selectivity of molecular recognition. J Proteome Res. 2007, 6: 611-620. 10.1021/pr060368gView ArticlePubMedGoogle Scholar
- Tseng GN, Sonawane KD, Korolkova YV, Zhang M, Liu J, Grishin EV, Guy HR: Probing the outer mouth structure of the HERG channel with peptide toxin footprinting and molecular modeling. Biophys J. 2007, 92: 3524-3540. 10.1529/biophysj.106.097360PubMed CentralView ArticlePubMedGoogle Scholar
- Korolkova YV, Kozlov SA, Lipkin AV, Pluzhnikov KA, Hadley JK, Filippov AK, Brown DA, Angelo K, Strobaek D, Jespersen T, Olesen SP, Jensen BS, Grishin EV: An ERG channel inhibitor from the scorpion Buthus eupeus. J Biol Chem. 2001, 276: 9868-9876. 10.1074/jbc.M005973200View ArticlePubMedGoogle Scholar
- Zeng XC, Luo F, Li WX: Molecular dissection of venom from Chinese scorpion Mesobuthus martensii: identification and characterization of four novel disulfide-bridged venom peptides. Peptides. 2006, 27: 1745-1754. 10.1016/j.peptides.2006.01.012View ArticlePubMedGoogle Scholar
- Peter M, Varga Z, Hajdu P, Gaspar R, Damjanovich S, Horjales E, Possani LD, Panyi G: Effects of toxins Pi2 and Pi3 on human T lymphocyte Kv1.3 channels: the role of Glu7 and Lys24. J Membr Biol. 2001, 179: 13-25. 10.1007/s002320010033View ArticlePubMedGoogle Scholar
- Han S, Yin SJ, Yi H, Mouhat S, Qiu S, Cao ZJ, Sabatier JM, Wu YL, Li WX: Protein-protein recognition control by modulating electrostatic interactions. J Proteome Res. 2010, 9: 3118-3125. 10.1021/pr100027kView ArticlePubMedGoogle Scholar
- Peigneur S, Beress L, Moller C, Mari F, Forssmann WG, Tytgat J: A natural point mutation changes both target selectivity and mechanism of action of sea anemone toxins. FASEB J. 2012, 26: 5141-5151. 10.1096/fj.12-218479View ArticlePubMedGoogle Scholar
- Korolkova YV, Bocharov EV, Angelo K, Maslennikov IV, Grinenko OV, Lipkin AV, Nosyreva ED, Pluzhnikov KA, Olesen SP, Arseniev AS, Grishin EV: New binding site on common molecular scaffold provides hERG channel specificity of scorpion toxin BeKm-1. J Biol Chem. 2002, 277: 43104-43109. 10.1074/jbc.M204083200View ArticlePubMedGoogle Scholar
- Jiang M, Zhang M, Maslennikov IV, Liu J, Wu DM, Korolkova YV, Arseniev AS, Grishin EV, Tseng GN: Dynamic conformational changes of extracellular S5-P linkers in the hERG channel. J Physiol. 2005, 569: 75-89. 10.1113/jphysiol.2005.093682PubMed CentralView ArticlePubMedGoogle Scholar
- Gilquin B, Braud S, Eriksson MA, Roux B, Bailey TD, Priest BT, Garcia ML, Menez A, Gasparini S: A variable residue in the pore of Kv1 channels is critical for the high affinity of blockers from sea anemones and scorpions. J Biol Chem. 2005, 280: 27093-27102. 10.1074/jbc.M413626200View ArticlePubMedGoogle Scholar
- Yin SJ, Jiang L, Yi H, Han S, Yang DW, Liu ML, Liu H, Cao ZJ, Wu YL, Li WX: Different residues in channel turret determining the selectivity of ADWX-1 inhibitor peptide between Kv1.1 and Kv1.3 channels. J Proteome Res. 2008, 7: 4890-48977. 10.1021/pr800494aView ArticlePubMedGoogle Scholar
- Han S, Yi H, Yin SJ, Chen ZY, Liu H, Cao ZJ, Wu YL, Li WX: Structural basis of a potent peptide inhibitor designed for Kv1.3 channel, a therapeutic target of autoimmune disease. J Biol Chem. 2008, 283: 19058-19065. 10.1074/jbc.M802054200View ArticlePubMedGoogle Scholar
- Liu J, Zhang M, Jiang M, Tseng GN: Structural and functional role of the extracellular s5-p linker in the HERG potassium channel. J Gen Physiol. 2002, 120: 723-737. 10.1085/jgp.20028687PubMed CentralView ArticlePubMedGoogle Scholar
- Visan V, Fajloun Z, Sabatier JM, Grissmer S: Mapping of maurotoxin binding sites on hKv1.2, hKv1.3, and hIKCa1 channels. Mol Pharmacol. 2004, 66: 1103-1112. 10.1124/mol.104.002774View ArticlePubMedGoogle Scholar
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