• 통합자연과학논문집
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• 제17권1호
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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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• 제32권5호
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• pp.459-468
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• 2007

이 연구에서는 법랑모세포 분화과정에서 APin의 기능을 알아보고자 APin-protein microarray를 시행한 후 치아발생과 관련이 있는 MEF2, Aurora kinase A, BMPR-IB와 EF-hand calcium binding protein을 분석하여 다음과 같은 결과를 얻었다. 1 CMV-APin construct를 transfection하여 APin의 과발현을 유도한 경우에는 MEF2와 Aurora kinase A 둘 모두에서 발현이 현저히 감소한 반면에, APin의 발현억제를 유도한 경우에는 둘 모두 변화가 없었다. 2. APin의 과발현을 유도한 경우에는 BMPR-IB와 EF-hand calcium binding protein 모두에서 발현이 크게 증가한 반면, APin을 발현억제 시킨 경우에는 BMPR-IB는 변화가 없었고, EF-hand calcium binding protein은 현저히 감소하였다. 위의 결과들로 보아 APin 단백질은 MEF2, Aurora kinase A, BMPR-IB, EF-hand calcium binding protein과 상호작용하여 법랑모세포의 분화와 석회화 과정 중에 중요한 역할을 하는 것으로 사료된다.

• 한국자원식물학회지
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• 제15권1호
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• pp.50-56
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• 2002

돈분뇨 발효액을 이용하여 녹색꽃양배추, 양미나리 관비재배 효과를 검정하고 이때의 적정 액비농도를 구명하고자 돈분뇨 발효액을 5, 10, 25 그리고 50배 희석(Ef. 5, Ef. 10, Ef. 25, Ef. 50)하여 시험한 결과는 다음과 같다. 작물 재배 후 토양분석결과 두 작물 공히 산도 및 칼륨은 액비농도가 진한 Ef. 5 처리구에서 낮았으나, Ef. 25, Ef. 50 처리구에서는 높았다. 반면 전기 전도도, 인산, 유기물함량, 그리고 질산태 질소는 액비농도가 낮은 Ef. 50 처리구에서 낮았으며, Ef. 5 처리구에서는 높아지는 경향이었다. 식물체 분석은 두작물 공히 총 질소는 액비농도가 진해짐에 따라 식물체내의 농도가 증가되었다. 유효 인산은 처리구에 관계없이 큰 차이가 없었다. 생육과 수량을 살펴보면 녹색꽃양배추의 경우 엽장, 엽폭 및 초장은 Ef. 50에서 생육이 가장 좋았다. 화뢰고와 화뢰폭은 액비농도가 낮은 Ef. 25와 Ef. 50 처리구에서 큰 것으로 나타났으며, 화뢰중 또한 무거웠다. 양미나리 는 엽장에서 Ef. 25와 Ef. 50 처리구에서 컸으며 ,수량은 관행대비 Ef. 25와 Ef. 50 처리구에서 각각 2배 증수되었으나, 고농도 처리인 Ef. 5에서 큰 폭으로 감소되었다. 이상의 결과 녹색꽃양배추 및 양미나리의 관비재배시 발효돈분뇨의 희석농도는 25∼50배가 적정한 것으로 나타났다.

• The Korean Journal of Physiology and Pharmacology
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• 제23권3호
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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.

• 스마트미디어저널
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• 제9권4호
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• pp.102-108
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• 2020

본 논문에서는 사람의 손동작에 의해 모바일장치상의 전기장센서를 통해 감지되는 동작신호의 실시간 검출 및 프레임 추출 알고리즘을 제안한다. 동작인식에 사용되는 전기장센서는 주변 환경 및 시점에 따라 랜덤잡음 및 센서 표면의 초기 대전상태의 가변적인 특성으로 인해 안정적으로 동작신호를 검출하는데 어려움이 있다. 본 논문에서는 이와 같은 환경에서도 안정적이고 강건하게 동작신호를 감지하여 검출할 수 있는 동적문턱치 방법(dynamic thresholding method)을 제안한다. 동작발생감지여부는 10Hz low-pass 필터와 MA(Motion Average) 필터를 통한 입력신호가 특정 문턱 전압값을 넘을 경우 감지되는데 감지 시점 센서상의 정전하상태가 가변적이므로 주기적으로 offset 값을 계산하여 새로운 문턱치를 동적으로 적용하는 방법이다. 이러한 방법으로 동작신호 감지율을 98% 이상으로 향상 시킬 수 있었다. 또한 일단 동작이 감지되면 정문턱치(positive thresold)와 부문턱치(negative threshold)의 통과시점, 횟수와 평균 동작주기를 고려한 동작신호프레임 알고리즘을 제안하였으며 이의 프레임추출 성공률도 98% 이상의 성능을 보였다. 본 논문에서 제안한 알고리즘으로 추출된 동작신호는 이후 신호정규화를 거쳐 LSTN 심층신경망 인식부를 거쳐 높은 손동작 인식률을 보임으로서 제안된 알고리즘의 우수함을 입증하였다.

• Biomolecules & Therapeutics
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• 제8권2호
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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.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 춘계총회 및 학술대회
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• pp.140.2-140.2
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• 2000

• Parasites, Hosts and Diseases
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• 제56권1호
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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.

• 상하수도학회지
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• 제22권5호
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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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• 제48권4호
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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.