• BMB Reports
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• 제52권3호
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• pp.163-164
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• 2019

The ribosomal synthesis of proteins in the eukaryotic cytosol has always been thought to start from the unformylated N-terminal (Nt) methionine (Met). In contrast, in virtually all nascent proteins in bacteria and eukaryotic organelles, such as mitochondria and chloroplasts, Nt-formyl-methionine (fMet) is the first building block of ribosomal synthesis. Through extensive approaches, including mass spectrometric analyses of the N-termini of proteins and molecular genetic techniques with an affinity-purified antibody for Nt-formylation, we investigated whether Nt-formylated proteins could also be produced and have their own metabolic fate in the cytosol of a eukaryote, such as yeast Saccharomyces cerevisiae. We discovered that Nt-formylated proteins could be generated in the cytosol by yeast mitochondrial formyltransferase (Fmt1). These Nt-formylated proteins were massively upregulated in the stationary phase or upon starvation for specific amino acids and were crucial for the adaptation to specific stresses. The stress-activated kinase Gcn2 was strictly required for the upregulation of Nt-formylated proteins by regulating the activity of Fmt1 and its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins could be distinct N-terminal degradation signals, termed fMet/N-degrons, and that Psh1 E3 ubiquitin ligase mediated the selective destruction of Nt-formylated proteins as the recognition component of a novel eukaryotic fMet/N-end rule pathway, termed fMet/N-recognin.

• 대한임상검사과학회지
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• 제52권3호
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• pp.172-180
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• 2020

본 연구의 목적은 병원정보시스템을 활용하여 N-말단 프로-뇌 나트륨 이뇨 펩타이드(N-terminal pro-brain natriuretic peptide, NT-proBNP)의 결과와 통상적인 혈액검사와의 상관성을 후향적으로 조사하는 것이다. NT-proBNP는 심부전의 병태 생리학에 관여한다. 본 연구결과는 연령과 NT-proBNP의 관계는 양의 상관관계(r=0.163)로 통계적으로 유의하였다(P<0.01). NT-proBNP 농도는 총 단백질(r=-0.250)과 알부민(r=-0.270)은 음의 상관관계, 적혈구수와 혈색소 그리고 적혈구 용적률은 음의 상관관계로 나타난 결과로 나타났다(P<0.01). NT-proBNP는 호중구와 양의 상관관계(r=0.227)와 림프구는 음의 상관관계(r=-0.236)로 통계적으로 유의한 결과가 나타났다(P<0.01). NT-proBNP와 크레아티닌은 양의 상관관계로 나타났으며(r=0.594, P<0.01), 다중회귀분석결과 가장 영향을 미치는 인자로 관찰되었다(B=0.53, t=7.65, P<0.01). NT-proBNP와 요산의 농도는 양의 상관관계로 유의한 관계를 나타냈다(r=0.180, P<0.05). 젖산탈수소효소가 NT-proBNP에 영향을 미치는 인자로 관찰되었다(B=0.20, t=3.28, P<0.01). 이러한 설명력은 43%의 영향력을 나타내어 NT-proBNP의 정확한 검사와 관련인자들은 심장표지자로서 임상적 가치가 있는 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제19권4호
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• pp.349-355
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• 2015

We examined the effects of peripherally or centrally administered botulinum neurotoxin type A (BoNT-A) on orofacial inflammatory pain to evaluate the antinociceptive effect of BoNT-A and its underlying mechanisms. The experiments were carried out on male Sprague-Dawley rats. Subcutaneous (3 U/kg) or intracisternal (0.3 or 1 U/kg) administration of BoNT-A significantly inhibited the formalin-induced nociceptive response in the second phase. Both subcutaneous (1 or 3 U/kg) and intracisternal (0.3 or 1 U/kg) injection of BoNT-A increased the latency of head withdrawal response in the complete Freund's adjuvant (CFA)-treated rats. Intracisternal administration of N-methyl-D-aspartate (NMDA) evoked nociceptive behavior via the activation of trigeminal neurons, which was attenuated by the subcutaneous or intracisternal injection of BoNT-A. Intracisternal injection of NMDA up-regulated c-Fos expression in the trigeminal neurons of the medullary dorsal horn. Subcutaneous (3 U/kg) or intracisternal (1 U/kg) administration of BoNT-A significantly reduced the number of c-Fos immunoreactive neurons in the NMDA-treated rats. These results suggest that the central antinociceptive effects the peripherally or centrally administered BoNT-A are mediated by transcytosed BoNT-A or direct inhibition of trigeminal neurons. Our data suggest that central targets of BoNT-A might provide a new therapeutic tool for the treatment of orofacial chronic pain conditions.

• Clinical and Experimental Pediatrics
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• 제52권2호
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• pp.234-241
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• 2009

목 적 : 가와사끼병에서 갑상샘 호르몬과 혈청 TNF-${\alpha}$, IL-6 및 NT-proBNP 농도와의 연관성을 알아보고자 하였다. 방 법 : 환자군으로 가와사끼병 환아 52명과 대조군으로 다른 열성 질환 환아 10명을 대상으로 하였다. 환자군의 치료 전 급성기와 치료 3-9일 후의 아급성기 때의 혈청과 대조군의 급성 발열기 혈청에서 갑상샘 호르몬과 TNF-${\alpha}$, IL-6, NT-proBNP를 각각 측정하여 비교하였다. 환자군에서는 심장 초음파검사로 관상동맥 병변의 유무를 조사하였다. 결 과 : 가와사끼병의 63.5%에서 저 $T_3$ 증후군($T_3$<100 ng/dL)이 동반되었다. 급성 가와사끼병의 $T_3$는 대조군에 비해 유의하게 낮았다($86.8{\pm}36.6$ vs $116.7{\pm}24.4$ ng/dL, P<0.05). 가와사끼병 환자군에서 $T_3$는 급성기에 감소하고 갑상샘 호르몬의 치료없이 아급성기에 증가하였으나($84.3{\pm}33.0$ vs $109.3{\pm}37.5$ ng/dL, P<0.01), 반대로 TNF-${\alpha}$, IL-6 및 NT-proBNP는 모두 급성기에 증가하고 아급성기에 감소하였다. 가와사끼병 환자군에서 저 $T_3$군(n=33)은 정상 $T_3$군(n=19)에 비해 신장 및 간기능의 차이는 없었고, NT-proBNP는 높았으며, IL-6는 약간 높은 경향을 보였다. 또한 $T_3$는 IL-6 (r=-0.12, P>0.05) 및 NT-proBNP(r=-0.54, P<0.001)와 반비례 관계를 보였다. 관상동맥 병변을 보인 가와사끼병 환아 4명 중 3명에서 $T_3$가 25.0-42.7 ng/dL로 매우 감소하였다. 면역글로불린 치료반응군(n=45)에 비해 저항군(n=7)에서 $T_3$는 더 낮았고($93.8{\pm}33.5$ vs $41.6{\pm}19.8$ ng/dL, P<0.001), IL-6와 NT-proBNP는 더 높았다. 결 론 : $T_3$는 가와사끼병의 급성기에 감소되며 아급성기에 갑상샘 호르몬의 치료 없이 정상화된다. $T_3$ 감소는 부분적으로 TNF-${\alpha}$ 보다는 IL-6의 작용에 의해 유발되며 NT-proBNP의 상승과 연관된다. 가와사끼병에서 $T_3$ 측정은 다른 열성 질환과의 감별진단에, 질병경과를 모니터하는데, 심근 손상의 초기 표식자로, 질병의 심한 정도를 예측하는데 이용할 수 있으리라 생각된다.

• Journal of Plant Biology
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• 제35권2호
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• pp.135-142
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• 1992

발아시킨 물오리나무(Alnus hirsuta)의 하배축 부위를 절단하여 얻은 떡잎이 포함된 상부를 6% glucose, MS(Murashige and Skoog, 1962) 비타민 혼합체, PVP(polyvinylpyrrolidone) 및 $0-50\;\mu\textrm{M}$이 BAP(6-benzylaminopurine)를 첨가한 NT(Nagata and Takebe, 1971) 무기염배지에서 배양한 결과, $5\;\mu\textrm{M}$이 BAP가 shoot 증식에 가장 적합한 것으로 나타났다. 따라서 이 배지에서 shoot 증식을 계속하여 실험에 사용할 200여개의 shoot를 확립하였다. Shoot 증식에 미치는 무기염류와 탄소원의 영향을 조사한 결과 탄소원에 무관하게 NT 무기염이 MS 무기염에 비해 3-12배의 증식을 나타냈으나 NT 무기염배지에서 탄소원은 특별한 차이를 보여주지 않았다. 발근에 미치는 NT 무기염, sucrose, IBA의 농도에 따른 영향을 조사한 결과 1/4-1/2 농도의 NT 무기염, 3%의 sucrose, $1-8\;\mu\textrm{M}$의 IBA에서 100% 가까운 발근율을 보였으며 뿌리발달도 양호하였다. 발근된 개체를 토양에 이식하고 습도를 점진적으로 낮추면서 적응시키면 거의 100%의 생존율을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.515-523
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• 2001

The dinoflagellates have some primitive nuclear features and are evolutionarily intermediate between prokaryotes and eukaryotes. The small subunit ribosomal RAN gene, the 5.8S ribosomal RNA gene, and the internal transcribed spacer (ITS) of Gonyaulax polyedra were cloned, and their sequences were analyzed to better understand their evolutionary position. The small subunit ribosomal RNA gene was 1,794 nt long, the large subunit ribosomal RNA gene was approximately 3,500 nt long, and the 5.8S ribosomal RNA gene was 159 nt long. The first internal transcribed spacer (ITS1) was 191 nt long, and the second internal transcribed spacer (ITS2) was 185 nt long. The intergenic spacer of the ribosomal RNA gene (IGS) was about 2,200 nt long, indicating that 5,800 nt of transcribed sequences were separated by roughly 2,200 nt of intergenic spacer. The ribosomal RNA genes were repeated many times and arranged in a head-to-tail, tandemly repeated manner. The repeating unit of ribosomal RNA gene of G. polyedra was proposed to be 8,000 nt long. Based on the lengths of ribosomal RNA, sequence alignments with representative organisms, and phylogenetic analysis on ribosomal RNA, G. polyedra appears to be one of the alveolates branched from the eukaryotic crown and, among dinoflagellates, it seems to not have emerged early.

• Molecules and Cells
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• 제45권3호
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• pp.158-167
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• 2022

Ubiquitin (Ub) is post-translationally modified by Ub itself or Ub-like proteins, phosphorylation, and acetylation, among others, which elicits a variety of Ub topologies and cellular functions. However, N-terminal (Nt) modifications of Ub remain unknown, except the linear head-to-tail ubiquitylation via Nt-Met. Here, using the yeast Saccharomyces cerevisiae and an Nt-arginylated Ub-specific antibody, we found that the detectable level of Ub undergoes Nt-Met excision, Nt-deamination, and Nt-arginylation. The resulting Nt-arginylated Ub and its conjugated proteins are upregulated in the stationary-growth phase or by oxidative stress. We further proved the existence of Nt-arginylated Ub in vivo and identified Nt-arginylated Ub-protein conjugates using stable isotope labeling by amino acids in cell culture (SILAC)-based tandem mass spectrometry. In silico structural modeling of Nt-arginylated Ub predicted that Nt-Arg flexibly protrudes from the surface of the Ub, thereby most likely providing a docking site for the factors that recognize it. Collectively, these results reveal unprecedented Nt-arginylated Ub and the pathway by which it is produced, which greatly expands the known complexity of the Ub code.

• Journal of Plant Biology
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• 제38권4호
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• pp.381-388
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• 1995

해녀콩(Canavalia lineata) 뿌리혹으로부터 만들어진 cDNA library를 뿌리의 총 RNA, leghemoglobin 및 uricasell cDNA를 competitor로 사용하여 차등 혼성화반응으로 후기 뿌리혹에서 발현되며 Cno이, Cne2, Cne3, Clb2로 명명된 4개의 cDNA 클론을 얻었다. Cnod1은 벼의 cysteine proteinase를 암호화하는 유전자와 유사한데 1450nt의 전사체외 혼성화 반응을 보였으며 뿌리혹에서만 발현되고 뿌리혹 발달 후기에서 가장 높은 발현을 보였다. Cne2는 벼의 Expressed Sequence Tag의 한 종류와 유사하고 900 nt의 전사체와 혼성화 반응을 보였으며 뿌리혹에서 주로 발현되었다. Cne2는 접종 후 13일째 발현이 증폭된 후 일정하게 유지되었다. Cne2는 완두의 tonoplast 막 내재 단백질 TRG31을 암호화하는 유전자와 유사하며 뿌리에서 가장 많이 발현되었다. Cne3는 1700 nt와 1400 nt의 전사체와 혼성화반응을 보였으며 뿌리혹의 성장, 발달과 일치하는 발현 양상을 보였다. Clb2는 800 nt의 전사체와 혼성화반응을 보였으며 접종 후 8일째부터 발현이 시작되어 13일째 증폭되고 그 이후 일정한 수준을 유지하였다.

• Molecules and Cells
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• 제19권3호
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• pp.328-333
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• 2005

Small heat shock proteins (sHSPs) are widely distributed, and their function and diversity of structure have been much studied in the field of molecular chaperones. In plants, which frequently have to cope with hostile environments, sHSPs are much more abundant and diverse than in other forms of life. In response to high temperature stress, sHSPs of more than twenty kinds can make up more than 1% of soluble plant proteins. We isolated a genomic clone, NtHSP18.3, from Nicotiana tabacum that encodes the complete open reading frame of a cytosolic class I small heat shock protein. To investigate the function of NtHSP18.3 in vitro, it was overproduced in Escherichia coli and purified. The purified NtHSP18.3 had typical molecular chaperone activity as it protected citrate synthase and luciferase from high temperature-induced aggregation. When E. coli celluar proteins were incubated with NtHSP18.3, a large proportion of the proteins remained soluble at temperatures as high as $70^{\circ}C$. Native gel analysis suggested that NtHSP18.3 is a dodecameric oligomer as the form present and showing molecular chaperone activity at the condition tested. Binding of bis-ANS to the oligomers of NtHSP18.3 indicated that exposure of their hydrophobic surfaces increased as the temperature was raised. Taken together, our data suggested that NtHSP18.3 is a molecular chaperone that functions as a dodecameric complex and possibly in a temperature-induced manner.

• 한국분말재료학회지
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• 제28권2호
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• pp.127-133
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• 2021

One-dimensional (1D) piezoelectric nanostructures are attractive candidates for energy generation because of their excellent piezoelectric properties attributed to their high aspect ratios and large surface areas. Vertically grown BaTiO₃ nanotube (NT) arrays on conducting substrates are intensively studied because they can be easily synthesized with excellent uniformity and anisotropic orientation. In this study, we demonstrate the synthesis of 1D BaTiO₃ NT arrays on a conductive Ti substrate by electrochemical anodization and sequential hydrothermal reactions. Subsequently, we explore the effect of hydrothermal reaction conditions on the piezoelectric energy conversion efficiency of the BaTiO₃ NT arrays. Vertically aligned TiO₂ NT arrays, which act as the initial template, are converted into BaTiO₃ NT arrays using hydrothermal reaction with various concentrations of the Ba source and reaction times. To validate the electrical output performance of the BaTiO₃ NT arrays, we measure the electricity generated from each NT array packaged with a conductive metal foil and epoxy under mechanical pushings. The generated output voltage signals from the BaTiO₃ NT arrays increase with increasing concentration of the Ba source and reaction time. These results provide a new strategy for fabricating advanced 1D piezoelectric nanostructures by demonstrating the correlation between hydrothermal reaction conditions and piezoelectric output performance.