scholarly journals Tryptophan 207 is crucial to the unique properties of the human voltage-gated proton channel, hHV1

2015 ◽  
Vol 146 (5) ◽  
pp. 343-356 ◽  
Author(s):  
Vladimir V. Cherny ◽  
Deri Morgan ◽  
Boris Musset ◽  
Gustavo Chaves ◽  
Susan M.E. Smith ◽  
...  
Keyword(s):  
Transmembrane Helix ◽  
Ph Sensor ◽  
Proton Channel ◽  
Tryptophan Residue ◽  
Ph Sensing ◽  
Ph Sensors ◽  
Voltage Gated ◽  
Karlodinium Veneficum ◽  
Channel Opening ◽  
External Ph

Part of the “signature sequence” that defines the voltage-gated proton channel (HV1) is a tryptophan residue adjacent to the second Arg in the S4 transmembrane helix: RxWRxxR, which is perfectly conserved in all high confidence HV1 genes. Replacing Trp207 in human HV1 (hHV1) with Ala, Ser, or Phe facilitated gating, accelerating channel opening by 100-fold, and closing by 30-fold. Mutant channels opened at more negative voltages than wild-type (WT) channels, indicating that in WT channels, Trp favors a closed state. The Arrhenius activation energy, Ea, for channel opening decreased to 22 kcal/mol from 30–38 kcal/mol for WT, confirming that Trp207 establishes the major energy barrier between closed and open hHV1. Cation–π interaction between Trp207 and Arg211 evidently latches the channel closed. Trp207 mutants lost proton selectivity at pHo >8.0. Finally, gating that depends on the transmembrane pH gradient (ΔpH-dependent gating), a universal feature of HV1 that is essential to its biological functions, was compromised. In the WT hHV1, ΔpH-dependent gating is shown to saturate above pHi or pHo 8, consistent with a single pH sensor with alternating access to internal and external solutions. However, saturation occurred independently of ΔpH, indicating the existence of distinct internal and external pH sensors. In Trp207 mutants, ΔpH-dependent gating saturated at lower pHo but not at lower pHi. That Trp207 mutation selectively alters pHo sensing further supports the existence of distinct internal and external pH sensors. Analogous mutations in HV1 from the unicellular species Karlodinium veneficum and Emiliania huxleyi produced generally similar consequences. Saturation of ΔpH-dependent gating occurred at the same pHo and pHi in HV1 of all three species, suggesting that the same or similar group(s) is involved in pH sensing. Therefore, Trp enables four characteristic properties: slow channel opening, highly temperature-dependent gating kinetics, proton selectivity, and ΔpH-dependent gating.

scholarly journals Histidine168 is crucial for ΔpH-dependent gating of the human voltage-gated proton channel, hHV1

2018 ◽  
Vol 150 (6) ◽  
pp. 851-862 ◽  
Author(s):  
Vladimir V. Cherny ◽  
Deri Morgan ◽  
Sarah Thomas ◽  
Susan M.E. Smith ◽  
Thomas E. DeCoursey
Keyword(s):  
Single Point ◽  
Species Difference ◽  
Transmembrane Helix ◽  
Proton Channel ◽  
Single Point Mutation ◽  
Ph Sensing ◽  
Electrophysiological Properties ◽  
Voltage Gated ◽  
Voltage Dependent ◽  
Kinetics Of

We recently identified a voltage-gated proton channel gene in the snail Helisoma trivolvis, HtHV1, and determined its electrophysiological properties. Consistent with early studies of proton currents in snail neurons, HtHV1 opens rapidly, but it unexpectedly exhibits uniquely defective sensitivity to intracellular pH (pHi). The H+ conductance (gH)-V relationship in the voltage-gated proton channel (HV1) from other species shifts 40 mV when either pHi or pHo (extracellular pH) is changed by 1 unit. This property, called ΔpH-dependent gating, is crucial to the functions of HV1 in many species and in numerous human tissues. The HtHV1 channel exhibits normal pHo dependence but anomalously weak pHi dependence. In this study, we show that a single point mutation in human hHV1—changing His168 to Gln168, the corresponding residue in HtHV1—compromises the pHi dependence of gating in the human channel so that it recapitulates the HtHV1 response. This location was previously identified as a contributor to the rapid gating kinetics of HV1 in Strongylocentrotus purpuratus. His168 mutation in human HV1 accelerates activation but accounts for only a fraction of the species difference. H168Q, H168S, or H168T mutants exhibit normal pHo dependence, but changing pHi shifts the gH-V relationship on average by <20 mV/unit. Thus, His168 is critical to pHi sensing in hHV1. His168, located at the inner end of the pore on the S3 transmembrane helix, is the first residue identified in HV1 that significantly impairs pH sensing when mutated. Because pHo dependence remains intact, the selective erosion of pHi dependence supports the idea that there are distinct internal and external pH sensors. Although His168 may itself be a pHi sensor, the converse mutation, Q229H, does not normalize the pHi sensitivity of the HtHV1 channel. We hypothesize that the imidazole group of His168 interacts with nearby Phe165 or other parts of hHV1 to transduce pHi into shifts of voltage-dependent gating.

scholarly journals Titanium Nitride Thin Film Based Low-Redox-Interference Potentiometric pH Sensing Electrodes

Sensors ◽  
2020 ◽  
Vol 21 (1) ◽  
pp. 42
Author(s):  
Shimrith Paul Shylendra ◽  
Wade Lonsdale ◽  
Magdalena Wajrak ◽  
Mohammad Nur-E-Alam ◽  
Kamal Alameh
Keyword(s):  
Thin Film ◽  
Titanium Nitride ◽  
Orange Juice ◽  
Ph Sensor ◽  
Reference Electrode ◽  
Cost Effective ◽  
Ph Sensing ◽  
Ph Sensors ◽  
Potential Shift ◽  
Double Junction

In this work, a solid-state potentiometric pH sensor is designed by incorporating a thin film of Radio Frequency Magnetron Sputtered (RFMS) Titanium Nitride (TiN) working electrode and a commercial Ag|AgCl|KCl double junction reference electrode. The sensor shows a linear pH slope of −59.1 mV/pH, R2 = 0.9997, a hysteresis as low as 1.2 mV, and drift below 3.9 mV/h. In addition, the redox interference performance of TiN electrodes is compared with that of Iridium Oxide (IrO2) counterparts. Experimental results show −32 mV potential shift (E0 value) in 1 mM ascorbic acid (reducing agent) for TiN electrodes, and this is significantly lower than the −114 mV potential shift of IrO2 electrodes with sub-Nernstian sensitivity. These results are most encouraging and pave the way towards the development of miniaturized, cost-effective, and robust pH sensors for difficult matrices, such as wine and fresh orange juice.

scholarly journals Ruthenium Oxide pH Sensing for Organs-On-Chip Studies

Proceedings ◽  
2018 ◽  
Vol 2 (13) ◽  
pp. 709 ◽  
Author(s):  
Esther Tanumihardja ◽  
Wouter Olthuis ◽  
Albert van den Berg
Keyword(s):  
Ph Sensor ◽  
Ruthenium Oxide ◽  
Open Circuit ◽  
Biological Medium ◽  
Similar Response ◽  
Ph Sensing ◽  
Ph Sensors ◽  
Cross Sensitivity ◽  
On Chip

A ruthenium oxide (RuOx) electrode is being developed as potentiometric pH sensor for organs-on-chip applications. Open-circuit potential (OCP) of the RuOx electrode showed a response of −58.05 mV/pH, with no cross-sensitivity to potentially interfering/complexing ions (tested were lithium, sulfate, chloride, and calcium ions). Similar response was observed in complex biological medium. The electrode stored in liquid had a long-term drift of −0.8 mV/hour (corresponding to ΔpH of 0.013/hour) and response time in complex biological medium was 3.7 s. Minimum cross-sensitivity to oxygen was observed as the OCP shifted ~3 mV going from deoxygenated to oxygenated solution. This response is one magnitude lower than previously reported for metal- oxide pH sensors. Overall, the RuOx pH sensor has proven to be a suitable pH sensor for organs- on-chip applications.

scholarly journals Hydrophobic gasket mutation produces gating pore currents in closed human voltage-gated proton channels

2019 ◽  
Vol 116 (38) ◽  
pp. 18951-18961 ◽  
Author(s):  
Richard Banh ◽  
Vladimir V. Cherny ◽  
Deri Morgan ◽  
Boris Musset ◽  
Sarah Thomas ◽  
...  
Keyword(s):  
Proton Channel ◽  
Voltage Gated ◽  
Closed State ◽  
Channel Opening ◽  
Proton Current ◽  
Voltage Dependent ◽  
Current Flows ◽  
Voltage Gated Ion Channels ◽  
Dynamics Simulations ◽  
Voltage Sensing Domain

The hydrophobic gasket (HG), a ring of hydrophobic amino acids in the voltage-sensing domain of most voltage-gated ion channels, forms a constriction between internal and external aqueous vestibules. Cationic Arg or Lys side chains lining the S4 helix move through this “gating pore” when the channel opens. S4 movement may occur during gating of the human voltage-gated proton channel, hHV1, but proton current flows through the same pore in open channels. Here, we replaced putative HG residues with less hydrophobic residues or acidic Asp. Substitution of individuals, pairs, or all 3 HG positions did not impair proton selectivity. Evidently, the HG does not act as a secondary selectivity filter. However, 2 unexpected functions of the HG in HV1 were discovered. Mutating HG residues independently accelerated channel opening and compromised the closed state. Mutants exhibited open–closed gating, but strikingly, at negative voltages where “normal” gating produces a nonconducting closed state, the channel leaked protons. Closed-channel proton current was smaller than open-channel current and was inhibited by 10 μM Zn2+. Extreme hyperpolarization produced a deeper closed state through a weakly voltage-dependent transition. We functionally identify the HG as Val109, Phe150, Val177, and Val178, which play a critical and exclusive role in preventing H+ influx through closed channels. Molecular dynamics simulations revealed enhanced mobility of Arg208 in mutants exhibiting H+ leak. Mutation of HG residues produces gating pore currents reminiscent of several channelopathies.

scholarly journals Laser-Induced Biochar Formation through 355 nm Pulsed Laser Irradiation of Wood, and Application to Eco-Friendly pH Sensors

Nanomaterials ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1904
Author(s):  
Sung-Yeob Jeong ◽  
Chan-Woo Lee ◽  
Jun-Uk Lee ◽  
Yong-Won Ma ◽  
Bo-Sung Shin
Keyword(s):  
Laser Irradiation ◽  
Agricultural Land ◽  
Ph Sensor ◽  
Pulsed Laser ◽  
Agricultural Products ◽  
Organic Polymer ◽  
Ph Sensing ◽  
Ph Sensors ◽  
Linear Change ◽  
Pulsed Laser Irradiation

Due to the limited availability of agricultural land, pH sensing is becoming more and more important these days to produce efficient agricultural products. Therefore, to fabricate eco-friendly and disposable sensors, the black carbon, which is called biochar, is formed by irradiation of a UV pulsed laser having a wavelength of 355 nm onto wood and applying the resulting material as a pH sensor. The surfaces of three types of wood (beech, cork oak, and ash) were converted to the graphitic structure after UV laser irradiation; their morphologies were investigated. In addition, since the content of lignin, an organic polymer, is different for each wood, optimal laser irradiation conditions (laser fluence) needed to form these woods into pH sensors were considered. Depending on the degree of oil-like material generated after laser irradiation, a disposable pH sensor that can be used from one to three times is fabricated; due to the environmental characteristics of wood and biochar, the sensor shows high availability in that it can be easily discarded after use on agricultural land. After that, it can be used as filter in soil. Our wood-based pH sensor sensitively measures sequential changes from pH 4 to pH 10 and shows a very linear change of △R/R, indicating its potential for use in agriculture.

scholarly journals TiO2 Nano Flowers Based EGFET Sensor for pH Sensing

Coatings ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 251
Author(s):  
Chih-Chiang Yang ◽  
Kuan-Yu Chen ◽  
Yan-Kuin Su
Keyword(s):  
Ph Value ◽  
Ph Sensor ◽  
High Sensitivity ◽  
Experimental Result ◽  
Ph Sensing ◽  
Voltage Reference ◽  
Ph Sensors ◽  
Oxide Substrate ◽  
Ph Range ◽  
The Relationship

In this study, pH sensors were successfully fabricated on a fluorine-doped tin oxide substrate and grown via hydrothermal methods for 8 h for pH sensing characteristics. The morphology was obtained by high-resolution scanning electron microscopy and showed randomly oriented flower-like nanostructures. The TiO2 nanoflower pH sensors were measured over a pH range of 2–12. Results showed a high sensitivity of the TiO2 nano-flowers pH sensor, 2.7 (μA)1/2/pH, and a linear relationship between IDS and pH (regression of 0.9991). The relationship between voltage reference and pH displayed a sensitivity of a 46 mV/pH and a linear regression of 0.9989. The experimental result indicated that a flower-like TiO2 nanostructure extended gate field effect transistor (EGFET) pH sensor effectively detected the pH value.

scholarly journals Voltage and pH difference across the membrane control the S4 voltage-sensor motion of the Hv1 proton channel

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
T. Moritz Schladt ◽  
Thomas K. Berger
Keyword(s):  
Membrane Potential ◽  
Patch Clamp ◽  
Conformational Changes ◽  
Voltage Sensor ◽  
Lung Epithelial Cells ◽  
Proton Channel ◽  
Ph Sensing ◽  
Voltage Gated ◽  
Lung Epithelial ◽  
The Difference

AbstractThe voltage-gated proton channel Hv1 is expressed in a variety of cells, including macrophages, sperm, and lung epithelial cells. Hv1 is gated by both the membrane potential and the difference between the intra- and extracellular pH (ΔpH). The coupling of voltage- and ∆pH-sensing is such that Hv1 opens only when the electrochemical proton gradient is outwardly directed. However, the molecular mechanism of this coupling is not known. Here, we investigate the coupling between voltage- and ΔpH-sensing of Ciona intestinalis proton channel (ciHv1) using patch-clamp fluorometry (PCF) and proton uncaging. We show that changes in ΔpH can induce conformational changes of the S4 voltage sensor. Our results are consistent with the idea that S4 can detect both voltage and ΔpH.

scholarly journals Identification of Extracellular Ph-Sensing Residues in the Voltage-Gated Proton Channel Hv1

2020 ◽  
Vol 118 (3) ◽  
pp. 266a
Author(s):  
Ashley L. Bennett ◽  
Guiliano Melki ◽  
I. Scott Ramsey
Keyword(s):  
Extracellular Ph ◽  
Proton Channel ◽  
Ph Sensing ◽  
Voltage Gated

scholarly journals Dimer interaction in the Hv1 proton channel

2020 ◽  
Vol 117 (34) ◽  
pp. 20898-20907
Author(s):  
Laetitia Mony ◽  
David Stroebel ◽  
Ehud Y. Isacoff
Keyword(s):  
Functional Analysis ◽  
Ion Channel ◽  
Coiled Coil ◽  
Voltage Sensor ◽  
Proton Channel ◽  
Dimer Interface ◽  
Voltage Sensors ◽  
Voltage Gated ◽  
Channel Opening ◽  
The Stability

The voltage-gated proton channel Hv1 is a member of the voltage-gated ion channel superfamily, which stands out in design: It is a dimer of two voltage-sensing domains (VSDs), each containing a pore pathway, a voltage sensor (S4), and a gate (S1) and forming its own ion channel. Opening of the two channels in the dimer is cooperative. Part of the cooperativity is due to association between coiled-coil domains that extend intracellularly from the S4s. Interactions between the transmembrane portions of the subunits may also contribute, but the nature of transmembrane packing is unclear. Using functional analysis of a mutagenesis scan, biochemistry, and modeling, we find that the subunits form a dimer interface along the entire length of S1, and also have intersubunit contacts between S1 and S4. These interactions exert a strong effect on gating, in particular on the stability of the open state. Our results suggest that gating in Hv1 is tuned by extensive VSD–VSD interactions between the gates and voltage sensors of the dimeric channel.

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