• Title/Summary/Keyword: protein structures

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Crystal Structure of SAV0927 and Its Functional Implications

  • Jeong, Soyeon;Kim, Hyo Jung;Ha, Nam-Chul;Kwon, Ae-Ran
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.500-505
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    • 2019
  • Staphylococcus aureus is a round-shaped, gram-positive bacterium that can cause numerous infectious diseases ranging from mild infections such as skin infections and food poisoning to life-threatening infections such as sepsis, endocarditis and toxic shock syndrome. Various antibiotic-resistant strains of S. aureus have frequently emerged, threatening human lives significantly. Despite much research on the genetics of S. aureus, many of its genes remain unknown functionally and structurally. To counteract its toxins and to prevent the antibiotic resistance of S. aureus, our understanding of S. aureus should be increased at the proteomic scale. SAV0927 was first sequenced in an antibiotic resistant S. aureus strain. The gene is a conserved hypothetical protein, and its homologues appear to be restricted to Firmicutes. In this study, we determined the crystal structure of SAV0927 at $2.5{\AA}$ resolution. The protein was primarily dimeric both in solution and in the crystals. The asymmetric unit contained five dimers that are stacked linearly with ${\sim}80^{\circ}$ rotation by each dimer, and these interactions further continued in the crystal packing, resulting in a long linear polymer. The crystal structures, together with the network analysis, provide functional implications for the SAV0927-mediated protein network.

Physiological activities of poly(amino acid)'s derivatives with β-sheet structure on the skin (베타시트 구조가 도입된 폴리아미노산 유도체의 피부활성에 관한 연구)

  • Shin, Sung Gyu;Han, Sa Ra;Jung, Naseul;Ji, Yoonsook;Jeong, Jae Hyun
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.6
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    • pp.1597-1604
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    • 2020
  • In this study, a synthesized poly(amino acid) self-assembly grafted with valine molecules was investigated on the skin activity of skin growth factors. The amphiphilic grafted poly(amino acid) derivatives were successfully synthesized by varying of degree of substitution(DS) and polymerization (DP) with valine molecules forming a β-sheet structure. Then, the pro-collagen biosynthesis of EGF(epidermal growth factor) was improved by 20%, and the inhibitory ability of tyrosinase activity was increased by 6.5 times by self-assembling of EGF with the poly(amino acid)s having β-sheet structures. This strategy of preparing protein self-assembly with poly(amino acid) derivatives will help improve the stability of protein growth factors and use it in medicals as well as cosmeceuticals through skin improvement.

Novel Anti-Angiogenic and Anti-Tumour Activities of the N-Terminal Domain of NOEY2 via Binding to VEGFR-2 in Ovarian Cancer

  • Rho, Seung Bae;Lee, Keun Woo;Lee, Seung-Hoon;Byun, Hyun Jung;Kim, Boh-Ram;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.29 no.5
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    • pp.506-518
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    • 2021
  • The imprinted tumour suppressor NOEY2 is downregulated in various cancer types, including ovarian cancers. Recent data suggest that NOEY2 plays an essential role in regulating the cell cycle, angiogenesis and autophagy in tumorigenesis. However, its detailed molecular function and mechanisms in ovarian tumours remain unclear. In this report, we initially demonstrated the inhibitory effect of NOEY2 on tumour growth by utilising a xenograft tumour model. NOEY2 attenuated the cell growth approximately fourfold and significantly reduced tumour vascularity. NOEY2 inhibited the phosphorylation of the signalling components downstream of phosphatidylinositol-3'-kinase (PI3K), including phosphoinositide-dependent protein kinase 1 (PDK-1), tuberous sclerosis complex 2 (TSC-2) and p70 ribosomal protein S6 kinase (p70S6K), during ovarian tumour progression via direct binding to vascular endothelial growth factor receptor-2 (VEGFR-2). Particularly, the N-terminal domain of NOEY2 (NOEY2-N) had a potent anti-angiogenic activity and dramatically downregulated VEGF and hypoxia-inducible factor-1α (HIF-1α), key regulators of angiogenesis. Since no X-ray or nuclear magnetic resonance structures is available for NOEY2, we constructed the three-dimensional structure of this protein via molecular modelling methods, such as homology modelling and molecular dynamic simulations. Thereby, Lys15 and Arg16 appeared as key residues in the N-terminal domain. We also found that NOEY2-N acts as a potent inhibitor of tumorigenesis and angiogenesis. These findings provide convincing evidence that NOEY2-N regulates endothelial cell function and angiogenesis by interrupting the VEGFR-2/PDK-1/GSK-3β signal transduction and thus strongly suggest that NOEY2-N might serve as a novel anti-tumour and anti-angiogenic agent against many diseases, including ovarian cancer.

On The Licensed Dealer in Landing Markets (중매인에 관한 연구)

  • 유충열
    • The Journal of Fisheries Business Administration
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    • v.5 no.1
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    • pp.9-42
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    • 1974
  • Korea is a peninsular country surrounded by seas on three sides, and the fishing industry has carried out the service of suppling to the people with important anima protein. The distribution of fishery products has very complicated structures, because the production is in charge of small producers scattering along the coast all over the country, while consumpstion is made by separate homes in areas away from producing district. The relation between these two factors the structures make very complicated. The most typical and special structures of fishery distridution are distinguished in two, that is, one is landing market, the other is inland market. Landing markets have been monopolized by fishermen's cooperatives, providing with landing facilities and building sites. Fish markets played not only an important role in the landing, but distribution and price determination of catches by auction or tender. Inland markets are two types of wholesale market in consuming center, one is the terminal market for urban consumers, the other type is the local market for rural consumers. Fundamental functions of landing markets are gathering, assessment, and distribution functions. Gathering function is in charge of wholesaler in fishery cooperative. Gathering amounts are equal to gathering capacity of wholesaler and transact ability of licensed dealers as shown below model. Gathering amount=f.gathering capacity(=pre price.landing facility.account of wholesaler.distance of fishing ground.conveniency purchasing.home port)=fㆍ transact ability of licensed dealers≒f.population or port, and table 1 indicates these relationship. Assessment and distribution functions are in charge of licensed dealers in consumption side. Assessment function should bring the value in production activity through the auction between the cooperative seller and the licensed dealer as buyer. For fair trade transaction in auction, the free competition is supposed to be a prerequisite among the licensed dealers. The ideal condition for free competition is sameness in the scale of buying amount by licensed delaers, but it is almost impossible to attain its goal in actual marketing.

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Structures of antimicrobial peptides HP (2-20) and interactions between HP(2-20) and membrain studied by NMR spectroscopy

  • Lee, Kwang-Hwan;Lee, Dong-Gun;Park, Yoonkyung;Hahm, Kyung-Soo;Kim, Yangmee
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.68-68
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    • 2003
  • HP(2-20) (AKKVFKRLEKLEKLFSKIQNDK) derived from the N-terminus of Helicobacter pylori Ribosomal Protein L1 shows potent antimicrobial activity against bacterial, fungi and cancer cells without cytotoxic effect. In order to investigate the relationships between antimicrobial activity and the structures, several analogues have been designed and synthesized. The structures of these peptides in SDS micelles have been investigated using NMR spectroscopy and they revealed that analogue 3 has the longest, well-defined alpha-helix from Val5 to Trp19. NOESY experiments performed on HP and its analogues in nondeuterated SDS micelles show that protons in the indole ring of Trp16 are in close contact with methylene protons of SDS micelles. In order to probe the position of HP and its analogues relative to the SDS micelles, spin-labeled stearate was added. Large effects are observed for the chemical shifts and the intensities of Phe5, Glu9, Phe12, and Trp16 within the helix region by 16-doxylstearate. This result implies that 16-doxylstearate is located in the center of the micelles and the hydrophobic phase of the amphiphilic ${\alpha}$-helix is located in contact with the acyl chains of the micelles. Also, Lys3 and Lys4 at N-terminus and Lys20 at C-terminus may produce an optimal arrangement for electrostatic interactions between the sulfate head groups of the SDS and the positively charged lysyl N$\sub$3/$\^$+/. Interactions between the indole ring of Trp and the membrane, as well as the amphiphilic ${\alpha}$-helical structure of HP induced by Trp at the C-terminus may allow HP to span the lipid bilayer. These structural features are crucial for their potent antibiotic activities.

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Correlation of Lignan Contents with Protein and Oil Contents in the Seeds of Sesamum indicum L.

  • Lee, Jin-Hwan;Baek, In-Youl;Ko, Jong-Min;Shim, Kang-Bo;Kang, Nam-Suk;Kim, Hyun-Tae;Kang, Churl-Hwan;Park, Keum-Yong;Park, Ki-Hun;Ha, Tae-Joung
    • Journal of Applied Biological Chemistry
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    • v.51 no.1
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    • pp.20-27
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    • 2008
  • The contents of lignan, protein, and oil of nine Sesamum indicum cultivars were analyzed. To determine the lignan contents, sesamin 1 and sesamolin 2 were isolated from the ethylacetate extracts of the Suwon cultivar, and the structures were fully characterized by spectral and physical methods. The seeds of nine cultivars were screened for two lignans, which were determined by HPLC using a $C_{18}$ reversed phase column coupled with a photodiode array detector. Suwon cultivar showed the highest concentration (1: $6.24{\pm}0.04$ and 2: $3.58{\pm}0.01\;mg/g$), whereas Soonheuk displayed the lowest (1: $0.91{\pm}0.01$ and 2: $0.73{\pm}0.01\;mg/g$). The average content ratio of sesamin 1 (3.64 mg/g) was significantly higher than that of sesamolin 2 (2.57 mg/g). The protein content ranged from $21.52{\pm}0.35$ to $31.22{\pm}0.25%$, Suwon containing the highest level and Soonheuk had the lowest. Kwangbaek showed the highest oil level ($49.84{\pm}0.40%$), while Soonheuk exhibited the lowest ($42.52{\pm}0.05%$). Sesamin 1 exhibited a stronger radical-scavenging activity in the ABTS ($IC_{50}:\;63.2{\pm}2.4\;{\mu}M$) than its DPPH radical-scavenging activity ($IC_{50}>200\;{\mu}M$). These results lead to the conclusion that lignan content is affected by protein and oil contents. Concentration of the chemical components in the cultivars could be a key factor in the selection process of a high quality species.

Characterization of Caveola-Vesicle Complexes (CVCs) Protein, PHIST/CVC-8195 in Plasmodium vivax

  • Wang, Bo;Lu, Feng;Han, Jin-Hee;Lee, Seong-Kyun;Cheng, Yang;Nyunt, Myat Htut;Ha, Kwon-Soo;Hong, Seok-Ho;Park, Won Sun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.725-732
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    • 2016
  • Plasmodium vivax produces numerous caveola-vesicle complex (CVC) structures beneath the membrane of infected erythrocytes. Recently, a member helical interspersed subtelomeric (PHIST) superfamily protein, $PcyPHIST/CVC-81_{95}$, was identified as CVCs-associated protein in Plasmodium cynomolgi and essential for survival of this parasite. Very little information has been documented to date about $PHIST/CVC-81_{95}$ protein in P. vivax. In this study, the recombinant $PvPHIST/CVC-81_{95}$ N and C termini were expressed, and immunoreactivity was assessed using confirmed vivax malaria patients sera by protein microarray. The subcellular localization of $PvPHIST/CVC-81_{95}$ N and C termini in blood stage parasites was also determined. The antigenicity of recombinant $PvPHIST/CVC-81_{95}$ N and C terminal proteins were analyzed by using serum samples from the Republic of Korea. The results showed that immunoreactivities to these proteins had 61% and 43% sensitivity and 96.9% and 93.8% specificity, respectively. The N terminal of $PvPHIST/CVC-81_{95}$ which contains transmembrane domain and export motif (PEXEL; RxLxE/Q/D) produced CVCs location throughout the erythrocytic-stage parasites. However, no fluorescence was detected with antibodies against C terminal fragment of $PvPHIST/CVC-81_{95}$. These results suggest that the $PvPHIST/CVC-81_{95}$ is localized on the CVCs and may be immunogenic in natural infection of P. vivax.

Domain Expression of ErmSF, MLS (macrolide-lincosamide-streptogramin B) Antibiotic Resistance Factor Protein (MLS (macrolide-lincosamide-streptogramin B) 항생제 내성인자 단백질인 ErmSF의 domain발현)

  • 진형종
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.245-252
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    • 2001
  • Erm proteins, MLS (macrolide-lincosamide-streptogramin B) resistance factor proteins, show high degree of amino acid sequence homology and comprise of a group of structurally homologous N-methyltransferases. On the basis of the recently determined structures of ErmC` and ErmAM, ErmSF was divided into two domains, N-terminal end catalytic domain and C-terminal end substrate binding domain and attempted to overexpress catalytic domain in E. coli using various pET expression systems. Three DNA fragments were used to express the catalytic domain: DNA fragment 1 encoding Met 1 through Glu 186, DNA fragment 2 encoding Arg 60 to Glu 186 and DNA fragment 3 encoding Arg 60 through Arg 240. Among the pET expression vectors used, pET 19b successfully expressed the DNA fragment 3 and pET23b succeeded in expression of DNA fragment 1 and 2. But the overexpressed catalytic domains existed as inclusion body, a insoluble aggregate. To assist the soluble expression of ErmSF catalytic domains, Coexpression of chaperone GroESL or Thioredoxin and lowering the incubation temperature to $22^{\circ}C$ were attempted, as did in the soluble expression of the whole ErmSF protein. Both strategies did not seem to be helpful. Solubilization with guanidine-HCl and renaturation with gradual removal of denaturant and partial digestion of overexpressed whole ErmSF protein (expressed to the level of 126 mg/ι culture as a soluble protein) with proteinase K, nonspecific proteinase are under way.

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Mechanosensitive Modulation of Receptor-Mediated Crossbridge Activation and Cytoskeletal Organization in Airway Smooth Muscle

  • Hai, Chi-Ming
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.535-547
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    • 2000
  • Recent findings indicate that mechanical strain (deformation) exerted by the extracellular matrix modulates activation of airway smooth muscle cells. Furthermore, cytoskeletal organization in airway smooth muscle appears to be dynamic, and subject to modulation by receptor activation and mechanical strain. Mechanosensitive modulation of crossbridge activation and cytoskeletal organization may represent intracellular feedback mechanisms that limit the shortening of airway smooth muscle during bronchoconstriction. Recent findings suggest that receptor-mediated signal transduction is the primary target of mechanosensitive modulation. Mechanical strain appears to regulate the number of functional G-proteins and/or phospholipase C enzymes in the cell membrane possibly by membrane trafficking and/or protein translocation. Dense plaques, membrane structures analogous to focal adhesions, appear to be the primary target of cytoskeletal regulation. Mechanical strain and receptor-binding appear to regulate the assembly and phosphorylation of dense plaque proteins in airway smooth muscle cells. Understanding these mechanisms may reveal new pharmacological targets for control1ing airway resistance in airway diseases.

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Reaction Mechanism of Vanadium Haloperoxidase and Marine Natural Products (Vanadium Haloperoxidase의 구조와 작용 메커니즘과 해양천연물질)

  • Han, Jae-Hong
    • Korean Journal of Crystallography
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    • v.16 no.2
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    • pp.66-74
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    • 2005
  • Marine natural products with various bioactivities are featured with similar structure to the common secondary metabolites and generally modified by halogenides, such as chloride, bromide, and iodide ions. Vanadium haloperoxidase is a key enzyme for the production of marine natural products and a metalloenzyme which requires a cofactor of vanadate. This review will cover isolation of vanadium haloperoxidase and the protein structures, as well as reaction mechanism of the metalloenzyme. Finally, reactivity of vanadium haloperoxidase and the biosynthesis of the secondary metabolites of indole, terpenoids, and acetogenins will be described.