• BMB Reports
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• 제54권3호
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• pp.157-163
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• 2021

The transient interactions between cellular components, particularly on membrane surfaces, are critical in the proper function of many biochemical reactions. For example, many signaling pathways involve dimerization, oligomerization, or other types of clustering of signaling proteins as a key step in the signaling cascade. However, it is often experimentally challenging to directly observe and characterize the molecular mechanisms such interactions-the greatest difficulty lies in the fact that living cells have an unknown number of background processes that may or may not participate in the molecular process of interest, and as a consequence, it is usually impossible to definitively correlate an observation to a well-defined cellular mechanism. One of the experimental methods that can quantitatively capture these interactions is through membrane reconstitution, whereby a lipid bilayer is fabricated to mimic the membrane environment, and the biological components of interest are systematically introduced, without unknown background processes. This configuration allows the extensive use of fluorescence techniques, particularly fluorescence fluctuation spectroscopy and single-molecule fluorescence microscopy. In this review, we describe how the equilibrium diffusion of two proteins, K-Ras4B and the PH domain of Bruton's tyrosine kinase (Btk), on fluid lipid membranes can be used to determine the kinetics of homodimerization reactions.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.186-196
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• 2023

Dyglomera^® is an aqueous ethanol extract derived from the fruit and pods of Dichrostachys glomerata. A previous study has revealed that Dyglomera regulates adipogenesis and lipolysis by modulating AMP-activated protein kinase (AMPK) phosphorylation and increased expression levels of lipolysis-related proteins in white adipose tissue of high fat diet-induced mice and 3T3-L1 adipocyte cells. To further investigate mechanisms of Dyglomera, additional studies were performed using 3T3-L1 cells. Results revealed that Dyglomera downregulated adipogenesis by inhibiting the protein kinase B/mammalian target of rapamycin signaling pathway and reconfirmed that it downregulated gene expression levels of proliferator-activated receptor (PPAR)-γ, CCAAT enhancer binding protein α, sterol-regulation element-binding protein-1c. Dyglomera also reduced adipokines such as tumor necrosis factor alpha, interleukin-1β, and interleukin 6 by regulating leptin expression. Moreover, Dyglomera promoted beige-and-brown adipocyte-related phenotypes and regulated metabolism by increasing mitochondrial number and expression levels of genes such as T-box protein 1, transmembrane protein 26, PR domain 16, and cluster of differentiation 40 as well as thermogenic factors such as uncoupling protein 1, proliferator-activated receptor-gamma co-activator-1α, Sirtuin 1, and PPARα through AMPK activation. Thus, Dyglomera not only can inhibit adipogenesis, but also can promote lipolysis and thermogenesis and regulate metabolism by affecting adipokine secretion from 3T3-L1 adipocytes.

• 대한화학회지
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• 제64권6호
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• pp.371-378
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• 2020

본 연구의 목적은 TIMSS 2019 중학교 화학 영역의 8개 주제를 우리나라 2009 개정 과학과 교육과정 및 2015 개정 과학과 교육과정에서 다루고 있는지 연계성을 분석하는 것이다. 이를 위해 초등학교 교사 4명과 중학교 교사 4명은 각각 물리, 화학, 생물, 지구과학 전공별로 참여하여 총 8명의 현장교사와 과학교육 전문가 2명이 참여해 우리나라 과학과 교육과정 중 TIMSS 2019 과학 평가틀의 화학 내용이 어느 학년에서 다루고 있는지를 분석하였다. 또한 본 연구는 TIMSS 2019 평가에서 8학년 대상의 246개 문항과 우리나라 과학과 교육과정 내용이 일치하는지 분석하여 8개 주제가 어느 학년에서 다루고 있는지도 분석에 반영하였다. 본 연구의 결과 및 논의는 다음과 같다. 첫째, TIMSS 2019 평가 주제 중에서 우리나라 중학교 교육과정에서 전혀 다루고 있지 않는 주제는 주기율표, 화학 반응에서 물질과 에너지, 화학결합에서 전자의 역할이었다. 따라서, '원소를 규칙에 따라 배열하는 주기율표'에 대한 주제는 우리나라 중학교 교육과정에 조기 도입할 필요가 있다. 둘째, '주변의 발열 반응이나 흡열 반응', 그리고 '반응 속도에 영향을 미치는 요인' 등에 관한 주제도 국제적인 흐름을 고려하여 우리나라 중학교 교육과정에 조기 도입하거나, 수준을 고려하면서 내용 등을 재구성하는 논의가 필요하다. 셋째, 차기 과학과 교육과정에서는 화학 내용과 범위, 특히 초·중학교 및 고등학교 과학과 교육과정에 포함될 개념의 범위와 순서가 연속적으로 연결되는 것 등과 관련된 시사점을 제공하였다.

• 공업화학
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• 제10권1호
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• pp.148-153
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• 1999

2-bromoterephthaloyl chloride와 1,4-bis(p-hydroxybenzoyloxy)butane을 PMMA 용액내에서 중축합시켜 in situ 복합재료를 제조하였다. 이들의 합성확인 및 열적, 기계적 성질은 FT-IR, FT-NMR, DSC와 DMTA에 의해서, 몰폴로지는 광학현미경과 SEM을 사용하여 조사하였다. 합성된 TLCP는 네마틱상을 보였고, 복합재료내 PMMA의 유리전이온도와 기계적 성질은 TLCP의 농도증가와 함께 증가하였다. TLCP domain은 PMMA 매트릭스내에서 미세 분산되었음을 확인하였고 in situ 중합에 의해 제조된 20 wt % TLCP/PMMA 복합재료는 같은 조성의 용액 블렌딩에 의해서 제조된 재료보다 더 높은 기계적 강도와 미세 분산도를 보였다.

• Bulletin of the Korean Chemical Society
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• 제28권9호
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• pp.1499-1509
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• 2007

Proteomic analyses of synaptic vesicle fraction from rat brain have been performed for the better understanding of vesicle regulation and signal transmission. Two different approaches were applied to identify proteins in synaptic vesicle fraction. First, the isolated synaptic vesicle proteins were treated with trypsin, and the resulting peptides were analyzed using a high-pressure capillary reversed phase liquid chromatography/tandem mass spectrometry (cRPLC/MS/MS). Alternatively, proteins were separated by two-dimensional gel electrophoresis (2DE) and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF/MS). Total 18 and 52 proteins were identified from cRPLC/MS-MS and 2DE-MALDI-TOF-MS analysis, respectively. Among them only 2 proteins were identified by both methods. Of the proteins identified, 70% were soluble proteins and 30% were membrane proteins. They were categorized by their functions in vesicle trafficking and biogenesis, energy metabolism, signal transduction, transport and unknown functions. Among them, 27 proteins were not previously reported as synaptic proteins. The cellular functions of unknown proteins were estimated from the analysis of domain structure, expression profile and predicted interaction partners.

• Journal of Microbiology and Biotechnology
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• 제32권3호
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• pp.287-293
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• 2022

The hydroxylation of methane (CH₄) is crucial to the field of environmental microbiology, owing to the heat capacity of methane, which is much higher than that of carbon dioxide (CO₂). Soluble methane monooxygenase (sMMO), a member of the bacterial multicomponent monooxygenase (BMM) superfamily, is essential for the hydroxylation of specific substrates, including hydroxylase (MMOH), regulatory component (MMOB), and reductase (MMOR). The diiron active site positioned in the MMOH α-subunit is reduced through the interaction of MMOR in the catalytic cycle. The electron transfer pathway, however, is not yet fully understood due to the absence of complex structures with reductases. A type II methanotroph, Methylosinus sporium 5, successfully expressed sMMO and hydroxylase, which were purified for the study of the mechanisms. Studies on the MMOH-MMOB interaction have demonstrated that Tyr76 and Trp78 induce hydrophobic interactions through π-π stacking. Structural analysis and sequencing of the ferredoxin domain in MMOR (MMOR-Fd) suggested that Tyr93 and Tyr95 could be key residues for electron transfer. Mutational studies of these residues have shown that the concentrations of flavin adenine dinucleotide (FAD) and iron ions are changed. The measurements of dissociation constants (K_ds) between hydroxylase and mutated reductases confirmed that the binding affinities were not significantly changed, although the specific enzyme activities were significantly reduced by MMOR-Y93A. This result shows that Tyr93 could be a crucial residue for the electron transfer route at the interface between hydroxylase and reductase.

• Korean Chemical Engineering Research
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• 제50권6호
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• pp.1068-1075
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• 2012

압출성형을 통한 core/shell 구조를 갖는 폴리부틸렌테레프탈레이트(PBT)/Nylon6,12 미세모를 제조함에 있어 최적의 압출조건을 규명하기 위하여, 압출온도와 배합비를 다르게 하여 제조한 블렌드 미세모의 상용성을 SEM 모폴로지와 DSC 분석을 통해 확인하고 UTM을 통해 압출속도에 따른 기계적 물성의 변화를 측정하였다. SEM 모폴로지 분석결과 압출온도가 증가할수록 분산상인 Nylon6,12 비드의 크기가 감소하였으며, Nylon6,12의 함량이 증가할수록 PBT 매트릭스 내 Nylon6,12의 상분리 현상이 감소하였다. DSC 분석 결과도 같은 경향을 나타냈는데, 압출온도가 상승함에 따라 녹는점에 해당하는 피크들의 경계가 사라지고, Nylon6,12의 비율이 증가할수록 두 피크의 간격이 좁아지는 것을 확인할 수 있었다. 한편 PBT/Nylon6,12 블렌드 미세모의 인장강도와 연신율 및 굴곡강도와 굴곡탄성률 모두 압출온도가 $260^{\circ}C$ 일 때까지 증가하였으나 그 이상의 온도에서는 오히려 감소하였다. $260^{\circ}C$에서의 인장강도와 연신율, 굴곡강도, 굴곡탄성률은 각각 560 $kg_f/cm^2$와 220%, 807 $kg_f/cm^2$, 22,146 $kg_f/cm^2$였는데 이는 PBT와 Nylon6,12의 중간 값을 상회하는 수치로 두 물질이 압출성형에 의한 블렌드 효과가 있다는 것을 확인할 수 있었다. 이처럼 우수한 상용성을 보일 때의 블렌드 압출 조건들을 토대로 하여 core/shell 구조의 이중구조 미세모를 제조하였다.

• Archives of Pharmacal Research
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• 제25권6호
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• pp.759-769
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• 2002

Mutated forms of ras are found in many human tumors and the rate of incidence is significantly higher in colon and pancreatic cancers. The protein product from the ras oncogene is a small G-protein, $p21^{ras}{\;}(Ras)$ that is known to playa key role in the signal transduction cascade and cell differentiation and proliferation. Mutated Ras is unable to regulate itself and remains constantly activated, leading to uncontrolled cell growth. The function of Ras in signal transduction requires its location near the growth factor receptor at the cell membrane. However, Ras does not have a transmembrane domain. Ras requires farnesylation to increase its hydrophobicity and subsequent plasma membrane association for its transforming activity. This key post-translational modification is catalyzed by the enzyme Ras farnesyltransferase (FTase), which transfers a farnesyl group from farnesylpyrophosphate to the C-terminal cysteine of the Ras protein. The requirement has focused attention on FTase as a target for therapeutic intervention. Selective inhibition of FTase will prevent Ras protein from association with the plasma membrane, leading to a disruption of oncogenic Ras function.

• Journal of Ginseng Research
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• 제21권3호
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• pp.147-152
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• 1997

Ginsenosides $Rh_1$ and $Rh_2$ Induced the differentiation of F9 teratocarcinoma stem cells. These agents are structurally similar to the steroid hormones, therefore, we speculated that the steroid receptor (s) or novel nuclear receptor (s) could be involved in the differentiation process induces by them. Based on this speculation, we tried to alone new nuclear receptors with reverse transcription-polymerase chain reaction (RT-PCR) method by isolating RNA from F9 teratocarcinoma cells induced by ginsenosides. By using RT-PCR with degenerated primers from highly conserved DNA binding domain of nuclear receptors, we identified several nuclear receptors. In northern blot analysis we found that these clones are transcriptionally regulated by ginsenoside Rhl or Rh2 treatment. Further characterizations of these clones are needed to identify the mechanism of gene expression, which has an important role in the differentiation of F9 cells induced by ginsenosides.

• 한국자기공명학회논문지
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• 제15권1호
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• pp.80-89
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• 2011

3D structures of STAM1 UIM-ubiquitin complex were presented to predict and analyze the interaction between UIM and ubiquitin. To generate the protein-peptide complex structure, the RosettaDock method was used with and without NMR restraints. High resolution complex structure was acquired successfully and evaluated electrostatic interaction in the protein-peptide binding with several charged residues at the binding site. From docking results, the Rosettadock method could be useful to acquire essential information of protein-protein or protein-peptide interaction with minimal biological evidences.