• Title/Summary/Keyword: proteome changes

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Proteome Analysis of Escherichia coli after High-dose Radiation

  • Lim, Sangyong;Lee, Misong;Joe, Minho;Song, Hyunpa;Kim, Dongho
    • Journal of Radiation Industry
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    • v.5 no.1
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    • pp.1-5
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    • 2011
  • Since proteomics can be employed to compare changes in the expression levels of many proteins under particular genetic and environmental conditions, using mass spectrometry to establish radiation stimulon, we performed two-dimensional gel electrophoresis and identified E. coli proteins whose expressions are affected by high dose of ionizing radiation. After exposure to 3 kGy, it was found that 6 proteins involved in carbon and energy metabolism were reduced. Although 4 of 7 protein spots showing a significant increase in expression level were neither identified nor classified, uridine phosphorylase (Udp), superoxide dismutase (SodB), and thioredoxin-dependent thiol peroxidase (Bcp) were proven to be up-regulated after irradiation. This suggests that E. coli subjected to high doses of radiation (3 kGy) may operate a defense system that is able to detoxify reactive oxygen species and stimulate the salvage pathway of nucleotide synthesis to replenish damaged DNA.

Proteomic Analysis of Serum Samples from Natural Herb and Immunoglobulin Yolksac (Ig Y) treated Porcine

  • Park, Hyeon-Soo;Nagappan, Arulkumar;Hong, Gyeong-Eun;Yumnam, Silvia;Lee, Ho-Jung;Kim, Gon-Sup
    • Journal of Korean Clinical Health Science
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    • v.1 no.1
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    • pp.83-89
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    • 2013
  • Purpose. Natural herb has been used for traditional medicine for thousands of years. For this reason, alternative medicine methods affect to domestic animal field. The main purpose of this study was to confirm the regulated proteins after feed additive treatment. Methods. We used total thirty male pigs were used for this experiment. E. coli and Salmonella typhimurium, were used for promote enteritis. Animals were divided into a negative control group, positive control group and test group to determine the effect of an additive mixture on the changes in protein expression in serum. The pigs were given a food supplemented with a natural herbal additive containing immunoglobulin yolksac (IgY) at concentrations 1%. On the 1st week and after eight weeks of feeding, the serums were collected from each group. The changes in the serum proteome as a response to the herbal additive were examined using two-dimensional polyacrylamide gel electrophoresis. Results. A total of 14 differentially expressed protein spots were identified by comparing the protein profiles of the control and additive treated porcine serums. Finally, 7 proteins were detected by MALDI-TOF/MS. Moreover, the proteins detected are involved in a cholesterol control factor, inflammation regulator, Conclusion. These results support of the hypothesis that a natural herbal additive containing IgY can affect the immune regulation system and reduce the pathological process by microbial infections.

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Protein Changes in Soymilk and Whole Soymilk due to Enzymatic Hydrolysis (효소적 가수분해에 따른 두유와 전두유의 단백질 변화)

  • Jang, Se-Young;Sin, Kyung-A;Park, Nan-Young;Bang, Kwang-Woong;Jeong, Yong-Jin
    • Food Science and Preservation
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    • v.15 no.6
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    • pp.903-908
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    • 2008
  • This study investigated protein changes in soymilk and whole soymilk due to enzymatic hydrolysis. The total free amino acid contents of low molecular weight soymilk (LSM) and low molecular weight whole soymilk (LWSM) were higher than soymilk (SM) and whole soymilk (WSM). The essential amino acid content was similar in SM and LSM, but was higher in LWSM than WSM. In SDS-PAGE performed to tendency of becoming low molecules, the soy protein molecular weights were 3372 kDa for SM and WSM, but 17 kDa or less for LSM and LWSM. Also, high molecular weight protein spots were evident in 2-D electrophoresis of SM and LSM, but only low molecular weight protein spots of various sizes were evident in WSM and LWSM. This suggests that the high molecular weight protein in SM and WSM is changed to low molecular weight protein by enzymatic hydrolysis. Further investigations of the separation and qualities of these proteins are required.

Proteome Changes in Penicillium expansum Grown in a Medium Derived from Host Plant

  • Xia, Xiaoshuang;Li, Huan;Liu, Fei;Zhang, Ye;Zhang, Qi;Wang, Yun;Li, Peiwu
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.624-632
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    • 2017
  • Penicillium expansum causes blue mold rot, a prevalent postharvest disease of pome fruit, and is also the main producer of the patulin. However, knowledge on the molecular mechanisms involved in this pathogen-host interaction remains largely unknown. In this work, a two-dimensional gel electrophoresis-based proteomic approach was applied to probe changes in P. expansum 3.3703 cultivated in apple juice medium, which was used to mimic the in planta condition. The results showed that the pH value and reducing sugar content in the apple juice medium decreased whereas the patulin content increased with the growing of P. expansum. A total of 28 protein spots that were up-regulated in P. expansum when grown in apple juice medium were identified. Functional categorization revealed that the identified proteins were mainly related to carbohydrate metabolism, secondary metabolism, protein biosynthesis or degradation, and redox homeostasis. Remarkably, several induced proteins, including glucose dehydrogenase, galactose oxidase, and FAD-binding monooxygenase, which might be responsible for the observed medium acidification and patulin production, were also detected. Overall, the experimental results provide a comprehensive interpretation of the physiological and proteomic responses of P. expansum to the host plant environment, and future functional characterization of the identified proteins will deepen our understanding of fungi-host interactions.

Comparative proteomic analysis of peripheral blood mononuclear cells from atopic dermatitis patients and healthy donors

  • Kim, Won-Kon;Cho, Hyun-Ju;Ryu, Su-In;Hwang, Hyang-Ran;Kim, Do-Hyung;Ryu, Hye-Young;Chung, Jin-Woong;Kim, Tae-Yoon;Park, Byoung-Chul;Bae, Kwang-Hee;Ko, Yong;Lee, Sang-Chul
    • BMB Reports
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    • v.41 no.8
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    • pp.597-603
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    • 2008
  • Atopic dermatitis (AD) is a chronic inflammatory skin disease that induces changes in various inflammatory skin cells. The prevalence of AD is as high as 18% in some regions of the world, and is steadily rising. However, the pathophysiology of AD is poorly understood. To identify the proteins involved in AD pathogenesis, a comparative proteomic analysis of protein expression in peripheral blood mononuclear cells isolated from AD patients and healthy donors was conducted. Significant changes were observed in the expressions of fourteen proteins, including the vinculin, PITPNB, and Filamin A proteins. Among the proteins, $\alpha$-SNAP and FLNA decreased significantly, and PITPNB increased significantly in AD patients compared with control subjects; these findings were further confirmed by real-time PCR and Western blot analysis. The comparative proteome data may provide a valuable clue to further understand AD pathogenesis, and several differentially regulated proteins may be used as biomarkers for diagnosis and as target proteins for the development of novel drugs.

Reduction of fetuin-A levels contributes to impairment of Purkinje cells in cerebella of patients with Parkinson's disease

  • Sunmi Yoon;Napissara Boonpraman;Chae Young Kim;Jong-Seok Moon;Sun Shin Yi
    • BMB Reports
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    • v.56 no.5
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    • pp.308-313
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    • 2023
  • Phenotypic features such as ataxia and loss of motor function, which are characteristics of Parkinson's disease (PD), are expected to be very closely related to cerebellum function. However, few studies have reported the function of the cerebellum. Since the cerebellum, like the cerebrum, is known to undergo functional and morphological changes due to neuroinflammatory processes, elucidating key functional factors that regulate neuroinflammation in the cerebellum can be a beneficial therapeutic approach. Therefore, we employed PD patients and MPTP-induced PD mouse model to find cytokines involved in cerebellar neuroinflammation in PD and to examine changes in cell function by regulating related genes. Along with the establishment of a PD mouse model, abnormal shapes such as arrangement and number of Purkinje cells in the cerebellum were confirmed based on histological finding, consistent with those of cerebellums of PD patients. As a result of proteome profiling for neuroinflammation using PD mouse cerebellar tissues, fetuin-A, a type of cytokine, was found to be significantly reduced in Purkinje cells. To further elucidate the function of fetuin-A, neurons isolated from cerebellums of embryos (E18) were treated with fetuin-A siRNA. We uncovered that not only the population of neuronal cells, but also their morphological appearances were significantly different. In this study, we found a functional gene called fetuin-A in the PD model's cerebellum, which was closely related to the role of cerebellar Purkinje cells of mouse and human PD. In conclusion, morphological abnormalities of Purkinje cells in PD mice and patients have a close relationship with a decrease of fetuin-A, suggesting that diagnosis and treatment of cerebellar functions of PD patients might be possible through regulation of fetuin-A.

Characterization of Placental Proteins in Bovine Somatic Cell Clone Fetuses

  • Woo, Jei-Hyun;Ko, Yeoung-Gyu;Kim, Bong-Ki;Kim, Jong-Mu;Lee, Youn-Su;Kim, Nam-Yun;Im, Gi-Sun;Yang, Boung-Chul;Seong, Hwan-Hoo;Jung, Jin-Kwan;Kwun, Moo-Sik;Chung, Hak-Jae
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.83-91
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    • 2005
  • Somatic cell nuclear transfer in cattle has limited efficiency in terms of production of live offspring due to high incidence of fetal failure after embryo transfer to recipients. Such low efficiency of cloning could possibly arise from abnormal and poorly developed placenta. In the present study the placental proteome in late pregnancy established from in vitro fertilization (IVF) and nuclear transfer (NT) was analysed. Proteome alternation was tested using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI- TOF). Comparing placenta from NT embryos to those from IVF counterparts, significant changes in expression level were found in 18 proteins. Of these proteins 12 were not expressed in NT placenta but expressed in IVF counterpart, whereas the expression of the other 6 proteins was limited only in NT placenta. Among these proteins, cytokeratin 8 and vimentin are considered to be involved in regulation of post-implantation development. In particular, cytokeratin 8 and vimentin may be used as makers for placental development during pregnancy because their expression levels changed considerably in NT placental tissue compared with its IVF counterpart. Data from 2-DE suggest that protein expression was disorientated in late pregnancy from NT, but this distortion was eliminated with progression of pregnancy. These findings demonstrate abnormal placental development during late pregnancy from NT and suggest that alterations of specific placental protein expression may be involved in abnormal function of placenta.

Proteomic Profiles of Mouse Neuro N2a Cells Infected with Variant Virulence f Rabies Viruses

  • Wang, Xiaohu;Zhang, Shoufeng;Sun, Chenglong;Yuan, Zi-Guo;Wu, Xianfu;Wang, Dongxia;Ding, Zhuang;Hu, Rongliang
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.366-373
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    • 2011
  • We characterized the proteomes of murine N2a cells following infection with three rabies virus (RV) strains, characterized by distinct virulence phenotypes (i.e., virulent BD06, fixed CVS-11, and attenuated SRV9 strains), and identified 35 changes to protein expression using two-dimensional gel electrophoresis in whole-cell lysates. The annotated functions of these proteins are involved in various cytoskeletal, signal transduction, stress response, and metabolic processes. Specifically, a-enolase, prx-4, vimentin, cytokine-induced apoptosis inhibitor 1 (CIAPIN1) and prx-6 were significantly up-regulated, whereas Trx like-1 and galectin-1 were down-regulated following infection of N2a cells with all three rabies virus strains. However, comparing expressions of all 35 proteins affected between BD06-, CVS-11-, and SRV9-infected cells, specific changes in expression were also observed. The up-regulation of vimentin, CIAPIN1, prx-4, and 14-3-3 ${\theta}/{\delta}$, and down-regulation of NDPK-B and HSP-1 with CVS and SRV9 infection were ${\geq}2$ times greater than with BD06. Meanwhile, Zfp12 protein, splicing factor, and arginine/serine-rich 1 were unaltered in the cells infected with BD06 and CVS-11, but were up-regulated in the group infected with SRV9. The proteomic alterations described here may suggest that these changes to protein expression correlate with the rabies virus' adaptability and virulence in N2a cells, and hence provides new clues as to the response of N2a host cells to rabies virus infections, and may also aid in uncovering new pathways in these cells that are involved in rabies infections. Further characterization of the functions of the affected proteins may contribute to our understanding of the mechanisms of RV infection and pathogenesis.

Proteomics of Liver Tissues of Bombina orientalis Following Exposure to Nonylphenol (Proteomics를 이용한 내분비계장애물질인 nonylphenol에 노출된 무당개구리의 단백질 발현 비교 연구)

  • Kim, Ho-Seung;Gye, Myung-Chan
    • Korean Journal of Ecology and Environment
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    • v.36 no.3 s.104
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    • pp.369-374
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    • 2003
  • Nonylphenol (NP), an well known aquatic contaminant, has been known to induce abnormalities in various aquatic animals. In an effort to develop proteome in the study of aquatic contamination of NP and its impact on the amphibia, protein changes in liver tissues of Korean red bellied frog, Bombina orientalis was investigated following the NP exposure. NP was administered intraperitoneally to male B. orientalis at 10 mg/kg body weight. At 48 and 96h after the treatment, the frog livers were sampled, and the protein fraction was separated using two dimensional gel electrophoresis (2D/E) and visualized with Coomassie brilluant blue staining. The 2D/E Images of the tissue from the animals treated with NP showed marked changes of protein spots (about 20% of total protein spots). Analysis of the 50-60 separated spots allowed identification of the major protein changes in the overall pattern for the stressor (NP) by time (0,48 and 96 h). At 48h after treatment, 8 spots were increased and 12 spots were reduced. Then, at 96h after treatment, 10 spots were increased and 8 spots were reduced. In total, approximately 29% of liver proteins showed the altered expression following the NP treatment. It is suggested that protein expression was repressed by blocking of certain metabolisms at 48 hand induced by the synthesis of new proteins for adaptation at 96 h following NP exposure. This application for 2D/E analysis may show promise in searching biomarkers for environmental proteomics in amphibians.

Expression of Heat Shock Protein and Antioxidant Genes in Rice Leaf Under Heat Stress

  • Lee, Dong-Gi;Ahsan, Nagib;Kim, Yong-Goo;Kim, Kyung-Hee;Lee, Sang-Hoon;Lee, Ki-Won;Rahman, Md. Atikur;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.3
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    • pp.159-166
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    • 2013
  • We have previously investigated the proteome changes of rice leaves under heat stress (Lee et al. in Proteomics 2007a, 7:3369-3383), wherein a group of antioxidant proteins and heat shock proteins (HSPs) were found to be regulated differently. The present study focuses on the biochemical changes and gene expression profiles of heat shock protein and antioxidant genes in rice leaves in response to heat stress ($42^{\circ}C$) during a wide range of exposure times. The results show that hydrogen peroxide and proline contents increased significantly, suggesting an oxidative burst and osmotic imbalance under heat stress. The mRNA levels of chaperone 60, HSP70, HSP100, chloroplastic HSP26, and mitochondrial small HSP responded rapidly and showed maximum expression after 0.5 or 2 h under heat stress. Transcript levels of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and Cu-Zn superoxide dismutase (Cu-Zn SOD) showed a rapid and marked accumulation upon heat stress. While prolonged exposure to heat stress resulted in increased transcript levels of monodehydroascorbate reductase, peroxidase, glyoxalase 1, glutathione reductase, thioredoxin peroxidase, 2-Cysteine peroxiredoxin, and nucleoside diphosphate kinase 1, while the transcription of catalase was suppressed. Consistent with their changes in gene expression, the enzyme activities of APX and DHAR also increased significantly following exposure to heat stress. These results suggest that oxidative stress is usually caused by heat stress, and plants apply complex HSP- and antioxidant-mediated defense mechanisms to cope with heat stress.