• Title/Summary/Keyword: component protein

검색결과 1,198건 처리시간 0.032초

Sensing the Stress: the Role of the Stress-activated p38/Hog1 MAPK Signalling Pathway in Human Pathogenic Fungus Cryptococcus neoformans

  • Bahn, Yong-Sun;Heitman, Joseph
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2007년도 International Meeting of the Microbiological Society of Korea
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    • pp.120-122
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    • 2007
  • All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.

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Streptococcus mutans GS5의 세포막 단백질 Ag I/II에 대한 단항체의 생산 (Generation of a monoclonal antibody against AgI/II, a cellular surface protein of Streptococcus mutans GS5)

  • 전철완;백병주;양연미;한지혜;김재곤
    • 대한소아치과학회지
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    • 제33권4호
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    • pp.587-596
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    • 2006
  • 치아우식증의 원인균으로 알려져 있는 Streptococcus mutans의 여러 균주 중 GS-5균주는 AgI/II 유전자 내에 생기는 nonsense mutation에 의하여 절편의 분비형 AgI/II 단백질을 발현한다. 이러한 사실은 S. nutans GS-5가 독특한 임상 기능을 가질 수 있음을 암시하며, 최근의 보고는 임상적으로 분리된 S. mutans 균주를 이용한 실험 결과에 근거하여 이러한 가능성을 지지한다. 본 연구는 이전의 실험을 통하여 S. mutans의 agI/II 유전자를 확보한 후 재조합 단백질인 N-terminal AgI/II 단백질을 생산하였다. 이 후 하이브리도마를 통하여 1C11A라 명명되는 세포막 단백질 AgI/II에 대한 단항체를 생산하였으며, Western blot과 ELISA를 통하여 이 항체가 AgI/II 단백질에 매우 높은 특이성을 나타냄을 알 수 있었다. 새로이 얻어진 단항체는 AgI/II와 관련된 질환의 기전을 규명하는데 기초 재료가 될 것이다.

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Effects of Ribosomal Protein L39-L on the Drug Resistance Mechanisms of Lung Cancer A549 Cells

  • Liu, Hong-Sheng;Tan, Wen-Bin;Yang, Ning;Yang, Yuan-Yuan;Cheng, Peng;Liu, Li-Juan;Wang, Wei-Jie;Zhu, Chang-Liang
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권7호
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    • pp.3093-3097
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    • 2014
  • Background: Cancer is a major threat to the public health whether in developed or in developing countries. As the most common primary malignant tumor, the morbidity and mortality rate of lung cancer continues to rise in recent ten years worldwide. Chemotherapy is one of the main methods in the treatment of lung cancer, but this is hampered by chemotherapy drug resistance, especially MDR. As a component of the 60S large ribosomal subunit, ribosomal protein L39-L gene was reported to be expressed specifically in the human testis and human cancer samples of various tissue origins. Materials and Methods: Total RNA of cultured drug-resistant and susceptible A549 cells was isolated, and real time quantitative RT-PCR were used to indicate the transcribe difference between amycin resistant and susceptible strain of A549 cells. Viability assay were used to show the amycin resistance difference in RPL39-L transfected A549 cell line than control vector and null-transfected A549 cell line. Results: The ribosomal protein L39-L transcription level was 8.2 times higher in drug-resistant human lung cancer A549 cell line than in susceptible A549 cell line by quantitative RT-PCR analysis. The ribosomal protein L39-L transfected cells showed enhanced drug resistance compared to plasmid vector-transfected or null-transfected cells as determined by methyl tritiated thymidine (3H-TdR) incorporation. Conclusions and Implications for Practice: The ribosomal protein L39-L gene may have effects on the drug resistance mechanism of lung cancer A549 cells.

Molecular Cloning and Functional Analysis of Rice (Oryza sativa L.) OsNDR1 on Defense Signaling Pathway

  • Lee, Joo-Hee;Kim, Sun-Hyung;Jung, Young-Ho;Kim, Jung-A;Lee, Mi-Ok;Choi, Pil-Gyu;Choi, Woo-Bong;Kim, Kyung-Nam;Jwa, Nam-Soo
    • The Plant Pathology Journal
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    • 제21권2호
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    • pp.149-157
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    • 2005
  • A novel rice (Oryza sativa L.) gene, homologous to Arabidopsis pathogenesis-related NDR1 gene, was cloned from cDNA library prepared from 30 min Magnaporthe grisea -treated rice seedling leaves, and named as OsNDR1. OsNDR1 encoded a 220-aminoacid polypeptide and was highly similar to the Arabidopsis AtNDR1 protein. OsNDR1 is a plasma membrane (PM)-localized protein, and presumes through sequence analysis and protein localization experiment. Overexpression of OsNDR1 promotes the expression of PBZ1 that is essential for the activation of defense/stressrelated gene. The OsNDR1 promoter did not respond significantly to treatments with either SA, PBZ, or ETP. Exogenously applied BTH induces the same set of SAR genes as biological induction, providing further evidence for BTH as a signal. Presumably, BTH is bound by a receptor and the binding triggers a signal transduction cascade that has an ultimate effect on transcription factors that regulate SAR gene expression. Thus OsNDR1 may act as a transducer of pathogen signals and/or interact with the pathogen and is indeed another important step in clarifying the component participating in the defense response pathways in rice.

산지에 따른 참깨종자의 이화학적 특성 비교 (The Comparison in the Physicochemical Properties of Sesame Seeds by Producing Areas)

  • 이민정;김기홍
    • Applied Biological Chemistry
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    • 제48권2호
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    • pp.128-131
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    • 2005
  • 국내산과 수입산 참깨의 이화학적 특성을 파악하기 위해 일반성분 분석, 지방산 조성 및 리그난 함량을 분석한 결과, 국내산, 수입산 참깨의 일반성분 분석결과 단백질함량은 국내산 참깨가 높았으며, 지방함량은 나이지리아산 참깨가 약간 높게 나타났다. 올레산과 리놀레산을 합한 총 양질 불포화지방산은 국내산 85.69%, 수입산 $83.06{\sim}85.05%$로 국내산 참깨가 양질의 지방산을 다량 함유하였다. 참깨 중 세사민과 세사몰린 성분 분석결과 국내산 참깨가 수입산 참깨보다 높은 함유량을 나타내었다. 참깨의 일반성분 분석결과 국내산, 수입산 참깨간에는 미미한 차이였지만 세사민과 세사몰린의 리그난 성분에서 국내산 참깨가 높은 수준으로 나타나 이화학적 특성이 우수한 것으로 나타났다.

Socioeconomic impact of traditional Korean medicine, Pyeongwee-San (KMP6) as an anti-allergic inflammatory drug

  • Song, Young-Hoon;Nam, Sun-Young;Choi, Young-Jin;Kim, Jeong-Hwa;Kim, Young-Sick;Jeong, Hyun-Ja
    • 셀메드
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    • 제2권3호
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    • pp.29.1-29.9
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    • 2012
  • The prevalence of allergic disease has been increasing over the past few decades in the majority of Western industrialized nations. There are some socioeconomic disparities regarding allergic disease status and management. Pyeongwee-San (KMP6) is Korean medicine for the treatment of gastrointestinal tract disease. It is known that KMP6 has an improving effect on the spleen and stomach functions in traditional Korean medical theory. Here, we hypothesized that KMP6 could be used to regulate the inflammatory reaction. We show the molecular mechanisms of Pyeongwee-San (KMP6) on inflammatory reactions. A molecular docking simulation showed that hesperidin, component of KMP6, regulate the enzymatic activity by interaction in the active site of caspase-1. KMP6 control the activity of caspase-1 in activated human mast cell line (HMC-1 cells). KMP6 reduced the expression of receptor interacting protein (RIP)-2 in HMC-1 cells. Thymic stromal lymphopoietin protein production and mRNA expression were inhibited by KMP6. In the activated HMC-1 cells, KMP6 suppressed the activation of mitogen-ativated protein kinase and nuclear factor-kappaB. In addition, KMP6 significantly inhibited the expression of inflammatory cytokines. Our findings indicate that KMP6 may attenuate allergic reactions via the regulation of caspase-1/RIP-2 signaling pathway. These studies will help advance the social welfare system.

스파르가눔 분비배설항원의 단백질 봉성 및 단백질분해효소 활성 (Component Proteins and Protease Activities in Excretory-Secretory Product of Sparganum)

  • 조승열;정영배;공윤
    • Parasites, Hosts and Diseases
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    • 제30권3호
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    • pp.227-230
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    • 1992
  • 스파르가눔증의 항체검사에서 항원으로 사용하는 스파르가눔 충체추출액의 구성 단백질 중, 환자혈청과 반응 시켜 관찰한 바, 분자량 36 및 31 kDa인 단백질이 항원성이 높았다. 또한 이 두가지 단백질은 스파르가눔 충체의 표피와 표피세포에 분포하여 충체 외부로 분비되는 항원일 것으로 추정하였다. 이 연구단보에서는 이들 두가지 단백질이 분비배설항원에 나타남을 증명하고자 스파르가눔을 $4^{\circ}C{\;}및{\;}37^{\circ}C$ 생리식염수에 30분~100시간 동안 배양하여 분비배설항원을 만들고, 분비배설항원내 단백질 분비량, 단백질의 조성 미치 단백질분해효소의 활성을 각각 관찰하였다. 충체 g당 분비배설항원의 단백질량은 관찰 범위안에서 배양온도에 관계없이 평균 7.7 mg이었다. SDS-폴리아크릴아미드 젤 전기영동상 분비배설항원의 단백질 구성은 분자량 66 kDa 이상인 단백질에서 차이가 있을 뿐 충체추출액의 것과 거의 유사하였다. 물론 분자량 36 및 31 kDa인 단백질도 분비배설항원의 주 구성성분이었다. 분비배설항원의 단백질분해효소 비활성도(比活性度)는 2.9~5.3 units/mg으로 충체추출액의 것과 차이가 없었다. 이미 보고된 스파르가눔의 cysteine protease가 체외로 분비되는 효소라는 것이 사실이라고 추정하게 하는 결과로 생각하였다.

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Characteristics of ustilago maydis virus of SH14 killer strain isolated in Korea

  • Hwang, Seon-Hee;Jung, Cheong-Hwan;Yie, Se-Won
    • Journal of Microbiology
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    • 제33권2호
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    • pp.154-159
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    • 1995
  • SH-14, a novel killer strain of Ustilago maydis was isolated in Korea. It has been reported in other papers that the toxin specificity and double-stranded RNA pattern of SH-14 strain were different from other laboratory strains. In this paper, we analyzed the biochemical characteristics of U. maydis SH-14 virus. Three distinctive peaks were isolated from CsCl density gradient, designated as top (T), intermediate (I) and bottom (B) components. We found that the densities of each components, 1.285, 1.408 g/cm$\^$3/, respectively, are very similar to those of other strains. As previously reported by the analysis of dsRNA in each component, the dsRNA segments are separately encapsidated. Capsid protein of SH-14 virus consists of two proteins about 70 Kd shown by SDS-PAGE analysis. Electron microscopic examination of the virus particles revealed that UmV particles are very similar in size and morphology to all isolates as well as all lab-strains. In order to test immunological cross reactivity of UmV, werstern bolt analysis was carriedout with antiserum against A8 virus. All capsid protein had positive reaction against A8 antibody which indicated that UmV are immunologically cross-reactive with all isolates from Korea. The results presented in this paper may show that UmV isolated from SH-14 strain has very similar biochemical characteristics to those of other UmV. However, the difference in the toxin specificity and the molecular weight of toxin protein from the SH-14 strain has us to conclude that U. maydis SH-14 strain is a new killer type.

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THE COMBINATION OF CHEMOMETRICS AND 2D NIR CORRELATION SPECTROSCOPY IN THE ANALYSIS OF DENATURATION PROCESS

  • Czarnik-Matusewicz, Boguslawa;Murayama, Koichi;Wu, Yuqing;Ozaki, Yukihiro
    • 한국근적외분광분석학회:학술대회논문집
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    • 한국근적외분광분석학회 2001년도 NIR-2001
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    • pp.1286-1286
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    • 2001
  • Despite extensive theoretical and experimental studies the structure of the protein-solvent interface is subject of many controversy. Understanding the processes that occur in aqueous solution requires understanding of the solvent influence on the structure of protein. The aim of this study is to investigate the applicability of NIR methods in the study of hydration phenomena in protein solutions. Temperature-induced changes in NIR spectra of -lactoglobulin (BLG) in aqueous solutions have been investigated by means of two-dimensional correlation spectroscopy (2DCOS) and principal component analysis (PCA). With the temperature increase the balance of forces between the BLG's interaction with itself and the BLGs interaction with its environment is disrupted leading to BLG unfolding. Significant differences of 2D signals and distinct discrepancies of loading on PC1 and PC2 were observed as a result of temperature increase. In the native folded conformation of BLC, most of the nonpolar amino acids are hidden in the centre of the structure, out of contact with water molecules, while charged groups are outside, in the contact with water. The polar groups promote low density Ih-type structure in the water outside this first hydration shell. When BLG unfolds it assumes a more extended configuration on which the previously buried nonpolar groups are exposed to water and promote the higher density II-type structure outside its first shell. Detailed assignments of bands attributed to the bulk water, different states of the hydrated water and the changed conformation of BLG are proposed.

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Effect of Different Tumbling Marination Methods and Time on the Water Status and Protein Properties of Prepared Pork Chops

  • Gao, Tian;Li, Jiaolong;Zhang, Lin;Jiang, Yun;Yin, Maowen;Liu, Yang;Gao, Feng;Zhou, Guanghong
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권7호
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    • pp.1020-1027
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    • 2015
  • The combined effect of tumbling marination methods (vacuum continuous tumbling marination, CT; vacuum intermittent tumbling marination, IT) and effective tumbling time (4, 6, 8, and 10 h) on the water status and protein properties of prepared pork chops was investigated. Results showed that regardless of tumbling time, CT method significantly decreased the muscle fiber diameter (MD) and significantly increased the total moisture content, product yield, salt soluble proteins (SSP) solubility, immobilized water component (p<0.05) compared with IT method. With the effective tumbling time increased from 4 h to 10 h, the fat content and the MD were significantly decreased (p<0.05), whereas the SSP solubility of prepared pork chops increased firstly and then decreased. Besides, an interactive effect between CT method and effective tumbling time was also observed for the chemical composition and proportion of immobilized water (p<0.05). These results demonstrated that CT method of 8 h was the most beneficial for improving the muscle structure and water distribution status, increasing the water-binding capacity and accelerating the marinade efficiency of pork chops; and thus, it should be chosen as the most optimal treatment method for the processing production of prepared pork chops.