• 제목/요약/키워드: PS I-mutant

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Synechocystis sp. PCC6803을 이용한 Photosystem I- mutants의 색소 및 틸라코이드막 단백질 분석 (Analysis of Pigments and Thylakoid Membrane Proteins in Photosystem I - Mutants from Synechocystis sp. PCC6803)

  • 전은경;장남기
    • 아시안잔디학회지
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    • 제11권1호
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    • pp.45-58
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    • 1997
  • Pigments and thylakoid membrane proteins were investigated in wild type and PS I- mutants from Synechocystis sp. PCC6803 Comparing morphological features, B2 was less fluorescent than the other strains. The contents of chlorophyll a were propotional to the FNR activity in thylakoid membrane. The FNR activity of mutants was lower than that of wild type. In the result of pigments analysis, mutants had smaller cholophyll a than that of wild type. The major carotenoid was found to he $\beta$-caroene, but aeaxanthin was barely detected in thylakoid membrane of mutants. The polypeptide, 14.8kD was detected by electrophoresis in mutants. It was considered to be the modification of 15.4kD in wild type. Membrane polypeptides of 17.6 and 19.7kD were not detected in mutants. In the result of western blotting, subunit I was detected in all strains, but subunit II was barely detected in mutants. Subunit II was not detected in B2 at all. In view of the results so far achieved, the changes of contents of chlorophyll and zeaxanthin were affected by the defficiency or modification of functional domain in subunit I. Also the modification in subunit I affected the subunit II- binding site in PS I. As the result, efficiency of photosynthesis was decreased. Key words: Synechoystis sp. PCC6803, PS I - mutant, Photosynthetic efficiency, Pigment,Thylakoid membrane proteins, Subunit I, II.

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인지질막 결합에 필요한 제5혈액응고인자 트립토판잔기들의 역할규명 (Characterization of tryptophan residues of human coagulation factor V required for binding to phospholipid membranes)

  • Kim, Suhng-Wook
    • 생명과학회지
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    • 제13권4호
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    • pp.463-472
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    • 2003
  • 제5인자와 지질막 phosphatidylserine과의 상호작용은 prothrombinase 복합체의 활성을 조절하는데 중요하다. 본 연구에서 제5인자의 지질 결합부위에 위치한 Trp2063과 Trp2064를 동시에 돌연변이 시킨 재조합 제5인자를 과발현 시키고 정제하였다. 돌연변이된 제5인자는 1-10%의 phosphatidylserine을 포함하는 지질막에서 아주낮은 활성을 보였다. surface plasmon resonance에 의해서 지질막과의 결합을 측정한 결과 돌연변이된 제5인자가 본래의 제5인자보다 고정된 지질막에의 결합이 현저하게 떨어지는 것을 관찰하였다. 제5인자가 phosphatidylserine을 포함하는 지질막에 높은 친화력으로 결합하기 위해서는 Trp2063과 Trp2064가 필수적이고 이러한 상호작용은 생리적인 phosphatidylserine 농도를 포함하는 지질막 위에서 prothrombinase 복합체의 형성에 필요하다는 결론을 내렸다.

Presenilin Modulates Calcium-permeant, Magnesium-Nucleotide regulated channel, I(MgNUM)

  • Shin, Sun-Young;Jeong, Soon-Youn;Uhm, Dae-Yong;Sungkwon Chung
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.47-47
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    • 2003
  • The presenilin 1 (PS1) or PS2 is an essential component of the ${\gamma}$-secretase complex, which mediates the intramembrane proteolysis of selected type-I membrane, including the ${\beta}$-amyloid precursor protein (APP) to yield A${\beta}$. Familial Alzheimer's disease (FAD)-associated mutations in presenilins give rise to an increased production of a highly amyloidogenic A${\beta}$42. In addition to their well-documented proteolytic function, the presenilins play a role in calcium signaling. We have previously reported that presenilin FAD mutations cause highly consistent alterations in intracellular calcium signaling pathways, which include deficits in capacitative calcium entry (CCE), the refilling mechanism for depleted internal calcium stores. However, molecular basis for the presenilin-mediated modulation of CCE remains to be elucidated. In the present study, whole-cell patch clamp method was used to identify a specific calcium-permeable ion channel current(s) that is responsible for the CCE deficits associated with FAD-linked PS1 mutants. Unexpectedly, both voltage-activated and conventional store depletion-activated calcium currents I(CRAC), were absent in HEK293 cells, which were stably transfected either with wild-type or FAD mutant (L286V, M146L, and delta E9) forms of PS1. Recently, magnesium-nucleotide-regulated metal cation current, or I(MagNum), has been described and appears to share many common properties with I(CRAC) including calcium permeability and inhibitor sensitivity (e.g. 2-APB). We have detected I(MagNum) in all 293 cells tested. Interestingly, FAD mutant 293 cells developed only about half of currents compared to PS1 wild type cells.

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A novel HDAC6 inhibitor, CKD-504, is effective in treating preclinical models of huntington's disease

  • Endan Li;Jiwoo Choi;Hye-Ri Sim;Jiyeon Kim;Jae Hyun Jun;Jangbeen Kyung;Nina Ha;Semi Kim;Keun Ho Ryu;Seung Soo Chung;Hyun Sook Kim;Sungsu Lee;Wongi Seol;Jihwan Song
    • BMB Reports
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    • 제56권3호
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    • pp.178-183
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    • 2023
  • Huntington's disease (HD) is a neurodegenerative disorder, of which pathogenesis is caused by a polyglutamine expansion in the amino-terminus of huntingtin gene that resulted in the aggregation of mutant HTT proteins. HD is characterized by progressive motor dysfunction, cognitive impairment and neuropsychiatric disturbances. Histone deacetylase 6 (HDAC6), a microtubule-associated deacetylase, has been shown to induce transport- and release-defect phenotypes in HD models, whilst treatment with HDAC6 inhibitors ameliorates the phenotypic effects of HD by increasing the levels of α-tubulin acetylation, as well as decreasing the accumulation of mutant huntingtin (mHTT) aggregates, suggesting HDAC6 inhibitor as a HD therapeutics. In this study, we employed in vitro neural stem cell (NSC) model and in vivo YAC128 transgenic (TG) mouse model of HD to test the effect of a novel HDAC6 selective inhibitor, CKD-504, developed by Chong Kun Dang (CKD Pharmaceutical Corp., Korea). We found that treatment of CKD-504 increased tubulin acetylation, microtubule stabilization, axonal transport, and the decrease of mutant huntingtin protein in vitro. From in vivo study, we observed CKD-504 improved the pathology of Huntington's disease: alleviated behavioral deficits, increased axonal transport and number of neurons, restored synaptic function in corticostriatal (CS) circuit, reduced mHTT accumulation, inflammation and tau hyperphosphorylation in YAC128 TG mouse model. These novel results highlight CKD-504 as a potential therapeutic strategy in HD.

Cyanobacterium Synechocystis sp.PCC6803 psaB 돌연변이주의 광합성 전자전달에 미치는 호흡의 영향 (Influence of Respiration on Photosynthetic Electron Transport in psaB Mutants from Cyanobacterium Synechocystis sp. PCC6803)

  • 윤병철;장남기
    • 아시안잔디학회지
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    • 제11권1호
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    • pp.59-72
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    • 1997
  • The influence of respiration on photosythetic electron transport were investigated in the Wid type and psaB mutants from Syneehocystis sp. PCC6803. The amount of glucose uptake in the wild type was proportional to the glucose concentration added in wild type and less than that of psaB mutants in the dark. It was suggested that psaB mutants more depend on the glucose than the wild type. It was investigated how the activities of isocitrate dehydrogenase(IDH) and glucose-6-phos-phate dehydrogenase(G6PDH) were changed. The activities of IDH were very low. While, the ac-tivities of G6PDH were much higher than that of IDH. These results agree to the reports that ex-ogenous glucose was dismilated aerobically via Oxidative Pentose Phosphate Pathway in heterotrophic cyanobacteria. PsaB mutants showed high G6PDH activity in the presence of glucose as well as in the dark and high respiratory activities especially in the dark. It was also investigated how photosynthetic electron transport activities were changed. PsaB mutants showed higher photosynthetic electron tranasport activities than wild type in the presence of glucose as well as in the dark. In the results, it was proposed that photosynthetic electron transport between PS I and PS U was complemented by respiratory electron transport through the NADPH generated by Dxidative Pentose Phophate Pathway in psaB mutant from Synechocystis sp. PCC6803. Key words: Photosynthetic electron transport, Respiration, Synechoystis sp. PCC6803, psaB mutant, Glucose uptake, IDH, G6PDH, Respiratory electron transport activity.

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New Classification of Plasmodiophora brassicae Races Using Differential Genotypes of Chinese Cabbage

  • Kim, Hun;Choi, Gyung Ja
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2015년도 춘계학술대회 및 임시총회
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    • pp.28-28
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    • 2015
  • Clubroot disease caused by Plasmodiophora brassicae induces severe losses of cruciferous vegetables worldwide. To control clubroot of Chinese cabbage, many CR (clubroot resistance) F1 hybrid cultivars have been bred and released in Korea, China and Japan. In this study, we determined the race of P. brassicae 12 field isolates, which collected from 10 regions in Korea, using Williams' differential varieties including two cabbage ('Jersey Queen', 'Badger Shipper') and two rutabaga ('Laurentian', 'Whilhelmsburger'). By Williams' differential varieties, 12 clubroot pathogens were assigned into one (GN2), two (HS and YC), two (HN1 and HN2), three (DJ, KS and SS) and four (GS, GN1, JS and PC) isolates for races 1, 2, 4, 5 and 9, respectively. In addition, the degree of resistance of 45 CR cultivars that were from Korea, China and Japan was tested with the 12 isolates. The 45 CR cultivars of Chinese cabbage were differentiated into three genotypes according to their resistance responses. Even though the 12 P. brassicae isolates were same race by Williams' differential varieties, three CR genotypes showed different resistance response to the isolates. These results indicate that races of P. brassicae by Williams' differentials were not related with resistance of CR cultivars, and three CR genotypes represented qualitative resistance to the P. brassicae isolates. CR genotype I including 'CR-Cheongrok' showed resistance to GN1, GN2, JS, GS, HS, DJ and KS isolates and susceptibility to YC, PC, HN1, HN2 and SS isolates. And CR genotype II such as 'Hangkunjongbyungdaebaekchae' was resistant to GN1, GN2, JS, GS, HS, YC, PC and HN1 and susceptible to DJ, KS, SS and HN2. CR genotype III including 'Chunhajangkun' and 'Akimeki' represented resistance to 10 isolates except for SS and HN2 isolates. Based on these results, we selected 'CR-Cheongrok', 'Hangkunjongbyungdaebaekchae', and 'Chunhajangkun' as a representative cultivar of three CR genotypes and 'Norangkimjang' as a susceptible cultivar. Furthermore, we investigated the resistance of 15 lines of Chinese cabbage, which were provided by seed companies, to 11 isolates except for HN1 of P. brassicae. The results showed that three lines were susceptible to all the tested isolates, whereas five, four, and three lines represented the similar responses corresponding to the CR genotypes I, II, and III, respectively; there is no line of Chinese cabbage showing different resistance patterns compared to three CR genotypes. In particular, line 'SS001' showing resistance responses of CR genotype II was a parent of 'Saerona' that have been commercialized as a CR $F_1$ cultivar of Chinese cabbage. Together, we divided 12 isolates of P. brassicae into 4 races, designated by wild type, mutant type 1, mutant type 2, and mutant type 3. Wild type including GN1, GN2, JS, GS, and HS isolates of P. brassicae was not able to infect all the cultivars of three CR genotypes, whereas, mutant type 3 such as SS and HN2 isolates developed severe clubroot disease on all the CR genotype cultivars. To mutant type 1 including DJ and KS isolates, CR genotypes I, II and III were resistant, susceptible and resistant, respectively. In contrast, to mutant type 2 including YC, PS, and HN1 isolates, CR genotypes I, II and III showed susceptibility, resistance and resistance, respectively. Taken together, our results provide the extended knowledge of classification of P. brassicae races, which is useful information for the breeding of resistant crops, with a suggestion that 'Norangkimjang', 'CR-Cheongrok', 'Saerona' and 'Chunhajangkun' cultivars of Chinese cabbage could be used as new race differentials of P. brassicae for clubroot disease assay.

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트레드밀 운동이 mutant (N141I) presenilin-2 유전자를 이식한 알츠하이머질환 모델 생쥐 뇌의 Aβ-42, cytochrome c, SOD-1, 2와 Sirt-3 단백질 발현에 미치는 영향 (The Effects of Treadmill Exercise on Cognitive Performance, Brain Mitochondrial Aβ-42, Cytochrome c, SOD-1, 2 and Sirt-3 Protein Expression in Mutant (N141I) Presenilin-2 Transgenic Mice of Alzheimer's Disease)

  • 구정훈;엄현섭;강은범;권인수;염동철;안길영;오유성;백영수;조인호;조준용
    • 생명과학회지
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    • 제20권3호
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    • pp.444-452
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    • 2010
  • 본 연구의 목적은 PS-2 (N141I) 알츠하이머 형질전환 모델 생쥐를 대상으로 트레드밀 운동이 뇌의 세포질과 미토콘드리아의 $A{\beta}$-42, cytochrome c, SOD-1, 2 and Sirt-3 단백질 발현에 미치는 효과를 알아보는데 있다. 우선 알츠하이머 형질전환 생쥐를 Non-Tg-sedentary (n=5), Non-Tg-treadmill exercise (n=5) 집단과 Tg-sedentary (n=5), Tg-treadmill exercise (n=5) 집단으로 구분하고 트레드밀 운동을 통한 신경보호 효과를 검증하기 위해 Tg와 Non-Tg집단에 12주간 트레드밀 운동을 수행한 후 인지능력을 살펴보고 뇌의 세포질과 미토콘드리아의 $A{\beta}$-42, cytochrome c, anti-oxidant enzymes (SOD-1, SOD-2)와 Sirt-3 단백질을 분석하였다. 먼저 트레드밀운동은 Tg 집단에서 인지능력의 개선을 나타냈으며 미토콘드리아의 $A{\beta}$-42와 세포질의 cytochrome c 단백질의 감소와 항산화 효소인 SOD-1, SOD-2를 유의하게 증가시켰다. 게다가 트레드밀 운동은 모든 집단에서 Sirt-3 단백질의 발현을 증가시켰다. 따라서 트레드밀 운동은 인지능력의 향상과 세포 내 스트레스를 유발하는 $A{\beta}$-42를 억제시켜 알츠하이머 질환을 개선시킬 수 있는 효과적인 방법이라고 생각된다.

종돈의 모근 Genomic DNA를 이용한 스트레스 증후군 검색 (Detection of Porcine Stress Syndrome from Genomic DNA of Hair Follicle by PCR-RFLP in Breeding Pig)

  • 김계웅;김진우;유재영;박홍양
    • Reproductive and Developmental Biology
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    • 제28권1호
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    • pp.37-43
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    • 2004
  • 본 연구는 319두의 서로 다른 품종에서 PSE육을 생산하는 PSS 돼지 출현빈도를 조사하였다(Yorkshire 150; Landrace 89 and Duroc 80). PCR-RFLP법을 이용하여 돼지의 모근을 DNA sample로 사용하여, PCR로 증폭된 유전자는 Cfo I 제한 효소로 절단하여 종돈에 존재하는 ryanodine receptor (RYR 1) 돌연변이 유전자의 출현빈도를 조사한 결과를 요약하면 다음과 같다. 모근에서 추출한 DNA를 주형으로 한 Primary PCR을 수행한 결과 ryanodine receptor 유전자 중 659bp의 증폭산물을 얻었으며, second PCR을 수행한 결과에서는 522 bp의 증폭산물을 얻었다. 이 증폭산물은 porcine ryanodine receptor 유전자의 exon 영역 중 PSS를 유발하는 point mutation(C\longrightarrowT; Arg\longrightarrowCys) 부분을 포함하고 있으므로 Cfo I 제한효소에 의해 분석될 수 있으며, agarose gel 전기영동에 의하여 세 가지의 유전자형으로 분류할 수 있다. 정상 homotype(NN)은 두 개의 DNA band(439, 83bp)로 나타나며, 열성 homotype(nn)은 552 bp의 단일 밴드로 출현한다. 그리고 세 개의 밴드(522, 439 그리고, 83 bp)는 heterotype(Nn)의 잠재성 돼지로 표현된다. Yorkshire종에서는 정상돼지가 98.00%로 나타났으며, hetero 돼지는 2.00% 그리고, PSS돼지는 출현하지 않았다. Landrace 돼지에서는 정상돼지가 87.64%로 나타났으며, hetero 돼지와 PSS패지가 각각 11.24와 1.12%로 나타났으나, Duroc종에서는 정장돼지(NN)만이 출현하였다. 대립 유전자 빈도는 Yorkshire종은 정상 N유전자가 0.990의 비율로 나타났으며, 열성 n 유전자는 0.010의 비율로 출현하였으며, Landrace종에서는 N유전자와 n유전자가 각각 0.933과 0.067의 빈도로 출현하였으며, Duroc종에서는 N 유전자의 빈도가 1.000의 빈도로 나타났으나, n유전자의 빈도는 0.000의 빈도로 나타났다. 3품종 집단 모두에서 Hardy-Weinberg 법칙과 일치하여 유전적 평형을 이루고 있었다.