• Journal of Integrative Natural Science
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• v.17 no.1
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• pp.31-41
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• 2024

Calcium ions play an important role in development and intracellular signaling. Dictyostelium discoideum has 14 genes encoding calcium -binding proteins (CBPs), but the function of most CBPs during development has not yet been studied. In this study, we investigated the specific functions of CBP7, one of 14 CBPs, in development using RNA interference cell lines of CBP7, cell lines overexpressing CBP7, cell lines with point mutations in the EF-hand domain, and cell lines expressing fragment proteins. was intended to reveal. CBP7 consists of 169 amino acids and contains 4EF-hand domains. The CBP7-overexpressing cells showed complete loss of developmental process. These cells remained in the single-cell growth stage under development -inducing conditions, while wild-type cells formed aggregations within 6-8h of development and eventually formed fruiting bodies. The experiments using point-mutated CBP7 protein showed that all EF-hand domains of CBP7 were important for CBP7 to function during developmental process. These results suggest that CBP7 plays an important role in developmental processes across all EF-hand domains.

• Restorative Dentistry and Endodontics
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• v.32 no.5
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• pp.459-468
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• 2007

Protein microarray or protein chips is potentially powerful tools for analysis of protein-protein interactions. APin cDNA was previously identified and cloned from a rat odontoblast cDNA library. The purpose of this study was to investigate the APin-protein interactions during ameloblast differentiation. Protein microarray was carried with recombinant APin protein and MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein were selected among 74 interacting proteins. Immortalized ameloblast cells (ALCs) were transfected with pCMV-APin construct and U6-APin siRNA construct. After transfection, the expression of the mRNAs for four proteins selected by protein micoarrays were assessed by RT-PCR. The results were as follows: 1. APin expression was increased and decreased markedly after its over-expression and inactivation, respectively. 2. Over-expression of the APin in the ALCs markedly down-regulated the expression of MEF2 and Aurora kinase A, whereas their expression remained unchanged by its inactivation. 3. Expression of BMPR-IB and EF-hand calcium binding protein were markedly increased by the over-expression of the APin in the ALCs, whereas expression of BMPR-IB remained unchanged and expression of EF-hand calcium binding protein was markedly decreased by its inactivation. These results suggest that APin plays an important role in ameloblast differentiation and mineralization by regulating the expression of MEF2, Aurora kinase A, BMPR-IB and EF-hand calcium binding protein.

• Korean Journal of Plant Resources
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• v.15 no.1
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• pp.50-56
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• 2002

This study was carried out to optimize the fertigation method using fermented swine liquid manure for the growth of two western vegetables, broccoli and celery. Plants were grown in a rain-shelter house and fertilized with a range of dilutions(efflux 5 dilution=Ef. 5, efflux 10 dilution=Ef. 10, efflux 25 dilution=Ef. 25, and efflux 50 dilution=Ef. 50) of the liquid manure or with conventional application of N : P$_2$O$\_$5/ : K$_2$O = 200 : 70 : 500kg/ha for broccoli, 250 : 210 : 240 kg/ha for celery as controls. After harvest, soil pH and K content decreased after using a high concentration of the liquid manure, Ef. 5, than after treatment with weaker concentrations at Ef. 25 and Ef. 50. On the other hand, soil electrical conductivity, content of P$_2$O$\_$5/, organic matter, total nitrogen, and NO$_3$-N at Ef. 5 increased as concentration of swine liquid manure increased. After harvest, available P$_2$O$\_$5/ in plant tissue did not differ significantly between any of the treatments. In broccoli, the lower concentration (Ef. 50) of swine liquid manure increased flowering over the other treatments, perhaps because the level of absorption into the plants is higher with lower concentration. The amounts of K and Ca in plant tissue were greatest after Ef. 25 and Ef. 50 treatments. Plant growth was best at Ef. 50 in broccoli, head height, head width, and head weight were the best with Ef. 25 and Ef. 50 treatments after harvest. In celery, leaf length was greater after Ef. 25 and Ef. 50 treatments than any other treatments. Total yield of celery of Ef. 25 and Ef. 50 treatments was twice that of conventional cultivation. On the other hand, yield severely decreased after application of high-concentration treatment at Ef. 5. In conclusion, fertigation of swine liquid manure, diluted in the range of Ef. 25 to Ef. 50, could improve yield and quality in broccoli and celery.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.3
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• pp.219-227
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• 2019

Polycystic kidney disease 2-like-1 (PKD2L1), polycystin-L or transient receptor potential polycystin 3 (TRPP3) is a TRP superfamily member. It is a calcium-permeable non-selective cation channel that regulates intracellular calcium concentration and thereby calcium signaling. Although the calmodulin (CaM) inhibitor, calmidazolium, is an activator of the PKD2L1 channel, the activating mechanism remains unclear. The purpose of this study is to clarify whether CaM takes part in the regulation of the PKD2L1 channel, and if so, how. With patch clamp techniques, we observed the current amplitudes of PKD2L1 significantly reduced when co-expressed with CaM and $CaM{\triangle}N$. This result suggests that the N-lobe of CaM carries a more crucial role in regulating PKD2L1 and guides us into our next question on the different functions of two lobes of CaM. We also identified the predicted CaM binding site, and generated deletion and truncation mutants. The mutants showed significant reduction in currents losing PKD2L1 current-voltage curve, suggesting that the C-terminal region from 590 to 600 is crucial for maintaining the functionality of the PKD2L1 channel. With PKD2L1608Stop mutant showing increased current amplitudes, we further examined the functional importance of EF-hand domain. Along with co-expression of CaM, ${\triangle}EF$-hand mutant also showed significant changes in current amplitudes and potentiation time. Our findings suggest that there is a constitutive inhibition of EF-hand and binding of CaM C-lobe on the channel in low calcium concentration. At higher calcium concentration, calcium ions occupy the N-lobe as well as the EF-hand domain, allowing the two to compete to bind to the channel.

• Smart Media Journal
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• v.9 no.4
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• pp.102-108
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• 2020

In this paper, we propose a real-time method of detecting hand motions and extracting the signal frame induced by EF(Electric Field) sensors. The signal induced by hand motion includes not only noises caused by various environmental sources as well as sensor's physical placement, but also different initial off-set conditions. Thus, it has been considered as a challenging problem to detect the motion signal and extract the motion frame automatically in real-time. In this study, we remove the PLN(Power Line Noise) using LPF with 10Hz cut-off and successively apply MA(Moving Average) filter to obtain clean and smooth input motion signals. To sense a hand motion, we use two thresholds(positive and negative thresholds) with offset value to detect a starting as well as an ending moment of the motion. Using this approach, we can achieve the correct motion detection rate over 98%. Once the final motion frame is determined, the motion signals are normalized to be used in next process of classification or recognition stage such as LSTN deep neural networks. Our experiment and analysis show that our proposed methods produce better than 98% performance in correct motion detection rate as well as in frame-matching rate.

• Biomolecules & Therapeutics
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• v.8 no.2
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• pp.153-159
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• 2000

A novel serine/threonine protein phosphatase with EF-hand motif, which belongs to PPEF family was partially cloned from rat brain cDNA by employing RT-PCR method. The size of the amplified clone was 1.6kbp. The amplified DNA was subcloned into pGEM-T-Easy vector and the resulting plasmid was maned as pGEM-rPPEF2. The nucleuotide sequence is shared by 88% with that of mouse PPEF-2 cDNA, and the deduced amino acid sequence reveal 92% homology with that of mouse PPEF-2 cDNA. The N-terminal region of the cloned rat brain PPEF contains a putative phosphatase catalytic domain (PP domain) and the C-terminal region contains multiple $Ca^{2+}$ binding sites (EF region). The putative catalytic domin (PP) and the EF-hand motif (EF) regions were subcloned into pGEX4T-1 and were overexpressed in E. coli DH5 as glutathione-S-transferase (GST) fusion proteins. Expression of the desired fusion protein was identified by SDS-PAGE and also by immunoblot analysis using monoclonal antibody against GST. The recombinant proteins were purified by glutathione-agarose chromatography. This report is first to demonstrate the cloning of PPEF family from rat brain tissues. The clone reported here would be invaluable for the investigation of the role of this new type-phosphatase in rat brain.

• Proceedings of the PSK Conference
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• 2000.04a
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• pp.140.2-140.2
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• 2000

• Parasites, Hosts and Diseases
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• v.56 no.1
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• pp.81-86
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• 2018

Four isoforms of calcium binding proteins containing 2 EF hand motifs and a dynein light chain-like domain in the human liver fluke Opisthorchis viverrini, namely OvCaBP1, 2, 3, and 4, were characterized. They had molecular weights of 22.7, 21.6, 23.7, and 22.5 kDa, respectively and showed 37.2-42.1% sequence identity to CaBP22.8 of O. viverrini. All were detected in 2- and 4-week-old immature and mature parasites. Additionally, OvCaBP4 was found in newly excysted juveniles. Polyclonal antibodies against each isoform were generated to detect the native proteins in parasite extracts by Western blot analysis. All OvCaBPs were detected in soluble and insoluble crude worm extracts and in the excretory-secretory product, at approximate sizes of 21-23 kDa. The ion-binding properties of the proteins were analyzed by mobility shift assays with the divalent cations $Ca^{2+}$, $Mg^{2+}$, $Zn^{2+}$, and $Cu^{2+}$. All OvCaBPs showed mobility shifts with $Ca^{2+}$ and $Zn^{2+}$. OvCaBP1 showed also positive results with $Mg^{2+}$ and $Cu^{2+}$. As tegumental proteins, OvCaBP1, 2, and 3 are interesting drug targets for the treatment of opisthorchiasis.

• Journal of Korean Society of Water and Wastewater
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• v.22 no.5
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• pp.551-559
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• 2008

An experimental study was carried out to evaluate the potential of flotation process for the secondary clarifier of an activated sludge process. Flotation techniques, applied in this study, include electrofloation (EF) which generated fine bubbles smaller than $35{\mu}m$ in average and diffuser flotation (DF) which generated fine bubbles smaller than $55{\mu}m$ in average. The batch experiments were done with activated sludge displaying various characteristics. It was shown that the efficiency of solids/liquid separation was reduced as the diluted sludge volume index ($DSVI_{30}$) of activated sludge increased. The dependency, however, gradually decreased as the gas to solids (G/S) ratio increased. Thickening efficiency of EF was more than 2~10 times and DF process was more than 1.5~5 times as compared with gravity sedimentation (GS). Stable sludge blanket was maintained regardless of sludge settleability when the G/S ratio was 0.019 in the EF. On the other hand, Serious deterioration in the sludge blanket was observed in the DF depends on G/S ratio and sludge settleability. And For EF and DF, the suspended solids concentration of effluent was not nearly influenced on settleability of activated sludge and more clear than GS. A biological nutrient removal (BNR) process, combined with EF as a secondary clarifier was operated for three months. The mean MLSS (mixed liquid suspended solids) concentration in the reactor and mean solids concentration of return sludge were estimated to be 5,340 mg/L and 16,770 mg/L, respectively. The water quality of effluent was considerably stable and low value was accomplished, that was, standard suspended solids concentration $0.07{\pm}0.51mg/L$ and standard turbidity $1.44{\pm}0.56NTU$. The EF could be applicable for enhancement of efficiency of activated sludge system as well as improvement of the water quality of effluent.

• Parasites, Hosts and Diseases
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• v.48 no.4
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• pp.319-324
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• 2010

A family of calcium-dependent protein kinases (CDPKs) is a unique enzyme which plays crucial roles in intracellular calcium signaling in plants, algae, and protozoa. CDPKs of malaria parasites are known to be key regulators for stage-specific cellular responses to calcium, a widespread secondary messenger that controls the progression of the parasite. In our study, we identified a gene encoding Plasmodium vivax CDPK4 (PvCDPK4) and characterized its molecular property and cellular localization. PvCDPK4 was a typical CDPK which had well-conserved N-terminal kinase domain and C-terminal calmodulin-like structure with 4-EF hand motifs for calcium-binding. The recombinant protein of EF hand domain of PvCDPK4 was expressed in Echerichia coli and a 34 kDa product was obtained. Immunofluorescence assay by confocal laser microscopy revealed that the protein was expressed at the mature schizont of P. vivax. The expression of PvCDPK4-EF in schizont suggests that it may participate in the proliferation or egress process in the life cycle of this parasite.