• BMB Reports
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• 제56권3호
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• pp.172-177
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• 2023

BEST family is a class of Ca²⁺-activated Cl- channels evolutionary well conserved from bacteria to human. The human BEST paralogs (BEST1-BEST4) share significant amino acid sequence homology in the N-terminal region, which forms the transmembrane helicases and contains the direct calcium-binding site, Ca²⁺-clasp. But the cytosolic C-terminal region is less conserved in the paralogs. Interestingly, this domain-specific sequence conservation is also found in the BEST1 orthologs. However, the functional role of the C-terminal region in the BEST channels is still poorly understood. Thus, we aimed to understand the functional role of the C-terminal region in the human and mouse BEST1 channels by using electrophysiological recordings. We found that the calcium-dependent activation of BEST1 channels can be modulated by the C-terminal region. The C-terminal deletion hBEST1 reduced the Ca²⁺-dependent current activation and the hBEST1-mBEST1 chimera showed a significantly reduced calcium sensitivity to hBEST1 in the HEK293 cells. And the C-terminal domain could regulate cellular expression and plasma membrane targeting of BEST1 channels. Our results can provide a basis for understanding the C-terminal roles in the structure-function of BEST family proteins.

• Environmental Engineering Research
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• 제21권4호
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• pp.341-349
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• 2016

To improve the utilization rate of construction waste as mine backfilling materials, this paper investigated the feasibility of using recycled powder as mine paste backfilling cementitious material, and studied the pozzolanic activity of recycled construction waste powder. In this study, alkali-calcium-sulfur served as the activation principle and an orthogonal test plan was performed to analyze the impact of the early strength agent, quick lime, and gypsum on the pozzolanic activity of the recycled powder. Our results indicated that in descending order, early strength agent > quick lime > gypsum affected the strength of the backfilling paste with recycled powder as a cementitious material during early phases. The strength during late phases was affected by, in descending order, quick lime > gypsum > early strength agent. Using setting time and early compressive strength as an analysis index as well as an extreme difference analysis, it was found that the optimal ratio of recycled powder cementitious material for mine paste backfilling was recycled powder:quick lime:gypsum:early strength agent at 78%:10%:8%:4%. X-ray diffraction analysis and scanning electron microscope were used to show that the hydration products of recycled powder cementitious material at the initial stages were mainly CH and ettringite. As hydration time increased, more and more recycled powder was activated. It mainly became calcium silicate hydrate, calcium aluminate hydrate, etc. In summary, recycled powder exhibited potential pozzolanic activities. When activated, it could replace cementitious materials to be used in mine backfill.

• The Korean Journal of Physiology and Pharmacology
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• 제21권2호
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• pp.241-249
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• 2017

Plasma membrane hyperpolarization associated with activation of $Ca^{2+}$-activated $K^+$ channels plays an important role in sperm capacitation during fertilization. Although Slo3 (slowpoke homologue 3), together with the auxiliary ${\gamma}^2$-subunit, LRRC52 (leucine-rich-repeat-containing 52), is known to mediate the pH-sensitive, sperm-specific $K^+$ current KSper in mice, the molecular identity of this channel in human sperm remains controversial. In this study, we tested the classical $BK_{Ca}$ activators, NS1619 and LDD175, on human Slo3, heterologously expressed in HEK293 cells together with its functional interacting ${\gamma}^2$ subunit, hLRRC52. As previously reported, Slo3 $K^+$ current was unaffected by iberiotoxin or 4-aminopyridine, but was inhibited by ~50% by 20 mM TEA. Extracellular alkalinization potentiated hSlo3 $K^+$ current, and internal alkalinization and $Ca^{2+}$ elevation induced a leftward shift its activation voltage. NS1619, which acts intracellularly to modulate hSlo1 gating, attenuated hSlo3 $K^+$ currents, whereas LDD175 increased this current and induced membrane potential hyperpolarization. LDD175-induced potentiation was not associated with a change in the half-activation voltage at different intracellular pHs (pH 7.3 and pH 8.0) in the absence of intracellular $Ca^{2+}$. In contrast, elevation of intracellular $Ca^{2+}$ dramatically enhanced the LDD175-induced leftward shift in the half-activation potential of hSlo3. Therefore, the mechanism of action does not involve pH-dependent modulation of hSlo3 gating; instead, LDD175 may modulate $Ca^{2+}$-dependent activation of hSlo3. Thus, LDD175 potentially activates native KSper and may induce membrane hyperpolarization-associated hyperactivation in human sperm.

• Advances in materials Research
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• 제4권3호
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• pp.133-144
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• 2015

The aim of this investigation is to prepare geopolymer resin by alkali activation of ceramic waste (AACW) with different sodium hydroxide (NaOH) and liquid sodium silicate (LSS) concentrations. In order to prepare geopolymer cement, AACW was replaced by 10 and 30 % by weight (wt.,) of concrete waste (CoW) as well as 10 and 30 wt., % ground granulated blast-furnace slag (GGBFS). The results showed that, the compressive strength of AACW increases with the increase of activator content up to 15:15 wt., % NaOH: LSS. All AACW hardened specimens activated by 3:3 (MC6), 6:6 (MC12), 12:12 (MC24) and 15:15 wt., % (MC30) NaOH: LSS destroyed when cured in water for 24h. The MC18 mix showed higher resistivity to water curing. The results also showed that, the replacement of AACW containing 9:9 wt., % NaOH: LSS (MC18) by 10 (MCCo10) and 30 (MCCo30) wt., % CoWdecreased the compressive strength at all ages of curing. In contrast, the MCCo10 mix showed the lower chemically combined water content compared to MC18 mix. The MCCo30 mix showed the higher chemically combined water content compared to MC18 and MCCo10 mixes. The compressive strength and chemically combined water of all AACWmixes containing GGBFS (MCS10 and MCS30) were higher than those of AACWwith no GGBFS (MC18). As the amount of GGBFS content increases the chemically combined water increases. The x-ray diffraction (XRD) proved that as the amount of CoWcontent increases, the degree of crystallinity increases. Conversely, the replacement of AACW by GGBFS leads to increase the amorphiticity character. The infrared spectroscopy (FTIR) confirms the higher reactivity of GGBFS compared to CoW as a result of successive hydration products formation, enhancing the compaction of microstructure as observed in scanning electron microscopy (SEM).

• 한국미생물·생명공학회지
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• 제14권4호
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• pp.285-291
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• 1986

효모Candida lipolytica 세포를 calcium alginate gel로 포괄 고정화시켜서. 유동층 반응기에서 반응을 수행하여 다음과 같은 결과를 얻었다. 1 고정화 효모 세포를 활성화 용액에서 회분식 유동층 반응기 방식과 연속식 유동층 반응기 방식으로 활성화시켰을 때, 세포는 고정화된 상태로 증식하였으며, 또한 세포당 시트르산 생성활성 이 증가하여서, 활성화되지 않은 bead보다 최대 시트르산 생성활성이 약 10배정도 증가되었다. 2 연속식 유동층 반응기 방식으로 활성화시킬 때가 회분식 유동층 반응기 방식으로 활성화시킬 때보다 늦은 시간에 최대의 시트르산 생성활성을 나타내었는데, 이것은 연속식으로 활성화시킬 때는 bead가 계속 새로운 환경에 놓이게 되어 bead내의 세포에 필요한 효소 및 보효소가 bead밖으로 계속 유출됨으로 인하여 bead내의 효소와 보효소의 축적에 많은 시간이 걸린데 기인한 것으로 사료된다. 3. 회분식 유동층 반응기내에서 세포수를 동일하게 하여 반응을 수행할 때, 고정화 bead의 크기가 작을수록 시트르산의 생산성이 증가하였다. 이것은 bead의 크기가 작을수록 부피에 비해 높은 표면적을 가지므로 세포의 많은 수가 반응에 참여하게 되며 bead내로의 화산저항이 작아서 물질전달이 잘되어 시트르산이 많이 생성된 것으로 사료된다.

• Molecules and Cells
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• 제44권2호
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• pp.88-100
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• 2021

Anoctamin 6/TMEM16F (ANO6) is a dual-function protein with Ca2+-activated ion channel and Ca2+-activated phospholipid scramblase activities, requiring a high intracellular Ca2+ concentration (e.g., half-maximal effective Ca2+ concentration [EC50] of [Ca2+]i > 10 μM), and strong and sustained depolarization above 0 mV. Structural comparison with Anoctamin 1/TMEM16A (ANO1), a canonical Ca2+-activated chloride channel exhibiting higher Ca2+ sensitivity (EC50 of 1 μM) than ANO6, suggested that a homologous Ca2+-transferring site in the N-terminal domain (Nt) might be responsible for the differential Ca2+ sensitivity and kinetics of activation between ANO6 and ANO1. To elucidate the role of the putative Ca2+-transferring reservoir in the Nt (Nt-CaRes), we constructed an ANO6-1-6 chimera in which Nt-CaRes was replaced with the corresponding domain of ANO1. ANO6-1-6 showed higher sensitivity to Ca2+ than ANO6. However, neither the speed of activation nor the voltage-dependence differed between ANO6 and ANO6-1-6. Molecular dynamics simulation revealed a reduced Ca2+ interaction with Nt-CaRes in ANO6 than ANO6-1-6. Moreover, mutations on potentially Ca2+-interacting acidic amino acids in ANO6 Nt-CaRes resulted in reduced Ca2+ sensitivity, implying direct interactions of Ca2+ with these residues. Based on these results, we cautiously suggest that the net charge of Nt-CaRes is responsible for the difference in Ca2+ sensitivity between ANO1 and ANO6.

• The Korean Journal of Physiology
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• 제27권2호
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• pp.175-183
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• 1993

There are many report suggesting that influx and intracellular calcium concentration $([Ca^{2+}]_i)$ are related to cell signalling in various cells. However, it has not been reported that calcium channel activation is affected by the substances involved in signal transduction pathways in the mouse eggs. In this study, the effects of isoprenaline (ISP) and cyclic AMP on calcium influx through calcium channels were investigated to show their relationship with the signal transduction process in unfertilized mouse eggs. Using whole cell voltage clamp techniques, calcium currents, elicited by the depolarizing pulses of 300 ms duration (from -50 mV to 50 mV in 10 mV increments) from a holding potential of -80 mV, were recorded. The current-voltage (I-V) relation of calcium currents was shown to be bell-shaped; the current began to activate at -50 mV and reached its maximum $(-1.33{\pm}0.16\;nA:\;mean{\pm}S.E.,\;n=7)$ at -10 mV, then decayed at around 50 mV. Calcium currents were fully activated within $7\;ms{\sim}20\;ms$ and completely inactivated 200 ms after onset of the step pulse. ISP within the concentration ranges of $10^{-8}\;M{\sim}10^{-4}\;M$ dose-dependently increased the amplitude calcium current. The permeable cyclic AMP analogue,8-bromocyclic AMP, also increased its maximal amplitude by 46ft at $10^{-5}\;M$, while protein kinase inhibitor (PKI), which is known to inhibit 0.02 phosphorylating units of cyclic AMP-dependent protein kinase (PKA) per microgram decreased calcium currents. Currents recorded in the presence of PKI were resistant to increase by the application of $10^{-5}\;M$. Also, PKI inhibited the calcium current increase elicited by ISP treatment. These results suggest that $\beta-adrenergic$ regulation of the calcium channel is mediated by the cAMP-dependent protein kinase. This signal transduction pathway might play a role in regulating $[Ca^{2+}]_i$, level due to the increase of calcium influx in mouse eggs.

• Biomolecules & Therapeutics
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• 제26권5호
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• pp.439-445
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• 2018

T-type calcium channels are low voltage-activated calcium channels that evoke small and transient calcium currents. Recently, T-type calcium channels have been implicated in neurodevelopmental disorders such as autism spectrum disorder and neural tube defects. However, their function during embryonic development is largely unknown. Here, we investigated the function and expression of T-type calcium channels in embryonic neural progenitor cells (NPCs). First, we compared the expression of T-type calcium channel subtypes (CaV3.1, 3.2, and 3.3) in NPCs and differentiated neural cells (neurons and astrocytes). We detected all subtypes in neurons but not in astrocytes. In NPCs, CaV3.1 was the dominant subtype, whereas CaV3.2 was weakly expressed, and CaV3.3 was not detected. Next, we determined CaV3.1 expression levels in the cortex during early brain development. Expression levels of CaV3.1 in the embryonic period were transiently decreased during the perinatal period and increased at postnatal day 11. We then pharmacologically blocked T-type calcium channels to determine the effects in neuronal cells. The blockade of T-type calcium channels reduced cell viability, and induced apoptotic cell death in NPCs but not in differentiated astrocytes. Furthermore, blocking T-type calcium channels rapidly reduced AKT-phosphorylation (Ser473) and $GSK3{\beta}$-phosphorylation (Ser9). Our results suggest that T-type calcium channels play essential roles in maintaining NPC viability, and T-type calcium channel blockers are toxic to embryonic neural cells, and may potentially be responsible for neurodevelopmental disorders.

• 한국축산식품학회지
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• 제21권4호
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• pp.285-291
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• 2001

본 연구는 활성탄을 사료에 첨가하여 그 첨가수준(0, 0.6, 0.9, 1.2%)에 따라 계육의 지방산 조성, 육색, 무김루의 변화를 검토하고자 broiler 48수를 공시하여 6주간 실험하였으며 특히 지방산과 무기물의 함량에 어떤 영향을 미치는가에 대하여 연구의 초점을 맞추었다. 대조구에 비해서 활성탄 0.6, 0.9%의 첨가구에서 oleic acid, arachidonic acid의 함량이 유의적으로 많았으며(p<0.05), 또한 활성탄 첨가수준에 따라 L*, a*, b* 값의 변화는 없었고, a* 값은 부위에 따라 유의성에 인정되었다(p<0.05). 활성탄 첨가수준에 따른 무기물 변화는 대조구에 비해 활성탄 첨가수준에 따른 무기물 변화는 대조구에 비해 활성탄 첨가구에서 Ca, Mg, P 함량이 많았으며(p<0.05) 총 무기물 함량은 활성탄 첨가구에서 많은 경향이었다. 따라서 본 연구의 결과 활성탄을 첨가하면 지방산은 oleic acid와 arachidonic acid 그리고 총무기물함량이 많아지는 경향이라고 결론 지을 수 있겠다.

• Journal of Korean Dental Science
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• 제10권2호
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• pp.45-52
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• 2017

Calcium has versatile roles in diverse physiological functions. Among these functions, intracellular $Ca^{2+}$ plays a key role during the secretion of salivary glands. In this review, we introduce the diverse cellular components involved in the saliva secretion and related dynamic intracellular $Ca^{2+}$ signals. Calcium acts as a critical second messenger for channel activation, protein translocation, and volume regulation, which are essential events for achieving the salivary secretion. In the secretory process, $Ca^{2+}$ activates $K^+$ and $Cl^-$ channels to transport water and electrolyte constituting whole saliva. We also focus on the $Ca^{2+}$ signals from intracellular stores with discussion about detailed molecular mechanism underlying the generation of characteristic $Ca^{2+}$ patterns. In particular, inositol triphosphate signal is a main trigger for inducing $Ca^{2+}$ signals required for the salivary gland functions. The biphasic response of inositol triphosphate receptor and $Ca^{2+}$ pumps generate a self-limiting pattern of $Ca^{2+}$ efflux, resulting in $Ca^{2+}$ oscillations. The regenerative $Ca^{2+}$ oscillations have been detected in salivary gland cells, but the exact mechanism and function of the signals need to be elucidated. In future, we expect that further investigations will be performed toward better understanding of the spatiotemporal role of $Ca^{2+}$ signals in regulating salivary secretion.