• Title/Summary/Keyword: Proteome analysis

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Proteome-based Identification of Proteins Regulated by Aldosterone in Rat Kidney Cortex (프로테오믹스 기술을 통한 쥐의 신장 피질에서 알도스테론(Aldosterone)에 의해 조절되는 단백질 동정)

  • Song, Mi-Na;Jeon, Hong-Bae;Choi, Hyo-Jung;Kwon, Tae-Hwan;Baek, Moon-Chang
    • YAKHAK HOEJI
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    • v.54 no.3
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    • pp.180-186
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    • 2010
  • Aldosterone, mineralocorticoid hormone, has important functions related to the regulation of blood pressure and balance of fluids and electrolytes in the distal region of the nephron. By genomic and non-genomic action of aldosterone, the physiological kidney functions are modulated. However, many of them except several kind of sodium channel have not been identified and analyzed yet. In this study, proteomic technologies with two-dimensional gel electrophoresis (2-DE) gel using aldosterone rat model were applied to analyze and identify the aldosterone dependently expressed proteins in rat kidney cortex. As a result, the established aldosterone rat model exhibited the normal physiological responses to aldosterone and modulated proteins were identified, which included 15 increased and 3 decreased proteins on 2-DE analysis. Among them, 11 proteins were identified as changed proteins by LC-MS/MS analysis. These proteins identified as aldosterone induced proteins were involved in several cellular pathways such as cytoskeleton remodeling, energy metabolism, amino acid metabolism, and chaperone process. In conclusion, our data could provide the insights into the new mechanism underlying regulation of kidney functions by aldosterone.

A Comprehensive Identification of Synaptic Vesicle Proteins in Rat Brains by cRPLC/MS-MS and 2DE/MALDI-TOF-MS

  • Lee, Won-Kyu;Kim, Hye-Jung;Min, Hye-Ki;Kang, Un-Beom;Lee, Cheol-Ju;Lee, Sang-Won;Kim, Ick-Young;Lee, Seung-Taek;Kwon, Oh-Seung;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.28 no.9
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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.

Anti-oxidant and Anti-aging Effect of Extracts from Seed of Camellia japonica L. by Supercritical Fluid Extraction (한국산 동백종자 초임계 추출물의 피부 항산화 및 항노화 연구)

  • Kim, Hee-Taek;Kim, Tae-Jun;Kim, Ee-Hwa;Kwak, Byeong-Mun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.33 no.4
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    • pp.32-43
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    • 2020
  • Objectives : The aim of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Camellia japonica L. extracted by supercritical CO2. Methods : The cell viability was performed by MTT assay. Nitric oxide (NO) production was performed by NO assay. The anti-oxidant effect was performed by beta-carotene bleaching assay and the intracellular proteome was analyzed expression of each 15 proteins by 2-D electrophoresis. Results : Fatty acid analysis of extract from Camellia japonica L. has shown oleic acid was 84%. And the antioxidant effect was about 186% compared with alpha-tocoperol(0.1%) by beta carotene bleaching assay. In 2D PAGE analysis, fifteen protein changes in five mechanisms which was collagen synthesis pathway, MMPs, ECM-cell interaction, cytokine, antioxidant enzymes were analyzed. Conclusions : On this study extract from seed of Camellia japonica L. by supercritical extraction could be used as a antioxidant and antiaging substance for the skin.

Integrated analysis of transcriptomic and proteomic analyses reveals different metabolic patterns in the livers of Tibetan and Yorkshire pigs

  • Duan, Mengqi;Wang, Zhenmei;Guo, Xinying;Wang, Kejun;Liu, Siyuan;Zhang, Bo;Shang, Peng
    • Animal Bioscience
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    • v.34 no.5
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    • pp.922-930
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    • 2021
  • Objective: Tibetan pigs, predominantly originating from the Tibetan Plateau, have been subjected to long-term natural selection in an extreme environment. To characterize the metabolic adaptations to hypoxic conditions, transcriptomic and proteomic expression patterns in the livers of Tibetan and Yorkshire pigs were compared. Methods: RNA and protein were extracted from liver tissue of Tibetan and Yorkshire pigs (n = 3, each). Differentially expressed genes and proteins were subjected to gene ontology and Kyoto encyclopedia of genes and genomes functional enrichment analyses. Results: In the RNA-Seq and isobaric tags for relative and absolute quantitation analyses, a total of 18,791 genes and 3,390 proteins were detected and compared. Of these, 273 and 257 differentially expressed genes and proteins were identified. Evidence from functional enrichment analysis showed that many genes were involved in metabolic processes. The combined transcriptomic and proteomic analyses revealed that small molecular biosynthesis, metabolic processes, and organic hydroxyl compound metabolic processes were the major processes operating differently in the two breeds. The important genes include retinol dehydrogenase 16, adenine phosphoribosyltransferase, prenylcysteine oxidase 1, sorbin and SH3 domain containing 2, ENSSSCG00000036224, perilipin 2, ladinin 1, kynurenine aminotransferase 1, and dimethylarginine dimethylaminohydrolase 1. Conclusion: The findings of this study provide novel insight into the high-altitude metabolic adaptation of Tibetan pigs.

Echinococcus granulosus Protoscolex DM9 Protein Shows High Potential for Serodiagnosis of Alveolar Echinococcosis

  • Kim, Jeong-Geun;Han, Xiumin;Kong, Yoon
    • Parasites, Hosts and Diseases
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    • v.60 no.1
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    • pp.25-34
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    • 2022
  • Alveolar echinococcosis (AE) caused by infection with E. multilocularis metacestode, represents one of the most fatal helminthic diseases. AE is principally manifested with infiltrative, proliferating hepatic mass, resembling primary hepatocellular carcinoma. Sometimes metastatic lesions are found in nearby or remote tissue. AE diagnosis largely depends on imaging studies, but atypical findings of imaging features frequently require differential diagnosis from other hepatic lesions. Serological tests may provide further evidence, while obtaining reliable AE materials is not easy. In this study, alternative antigens, specific to AE were identified by analyzing E. granulosus protoscolex proteins. An immunoblot analysis of E. granulosus protoscolex showed that a group of low-molecular-weight proteins in the range from 14 kDa to 16 kDa exhibited a sensitive and specific immune response to AE patient sera. Partial purification and proteomic analysis indicated that this protein group contained myosin, tubulin polymerization promoting protein, fatty-acid binding protein, uncharacterized DM9, heat shock protein 90 cochaperone tebp P-23, and antigen S. When the serological applicability of recombinant forms of these proteins was assessed using enzyme-linked immunosorbent assay, DM9 protein (rEgDM9) showed 90.1% sensitivity (73/81 sera tested) and 94.5% specificity (172/181 sera tested), respectively. rEgDM9 showed weak cross-reactions with patient sera from the transitional and chronic stages of cystic echinococcosis (3 to 5 stages). rEgDM9 would serve as a useful alternative antigen for serodiagnosis of both early- and advanced-stage AE cases.

Proteomic Analysis of Diesel Oil Biodegradation by Bacillus sp. with High Phosphorus Removal Capacity Isolated from Industrial Wastewater

  • Hee-Jung Kim;Deok-Won Kim;Jin-Hyeok Moon;Ji-Su Park;Eun-Ji Oh;Jin Yoo;Deok-Hyun Kim;Sun-Hwa Park;Keun-Yook Chung
    • Applied Chemistry for Engineering
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    • v.34 no.6
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    • pp.649-659
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    • 2023
  • This study was initiated to evaluate the phosphorus (P) removal and diesel oil degradation by bacteria isolated from industrial wastewater. The bacteria isolated were identified as Bacillus sp. The P removal efficiencies by Bacillus sp. were 99% at the initial 20 mg/L P concentration. The diesel degradation efficiencies by Bacillus sp. were 86.4% at an initial 1% diesel concentration. Lipophilicity by bacteria was the highest in the log phase, whereas it was the lowest in the death phase. As the diesel was used as a carbon source, P removal efficiencies by Bacillus sp. were 68%. When glucose, acetate, and a mixture of glucose and acetate as second carbon sources were added, the diesel degradation efficiencies were 69.22%, 65.46%, and 51.46%, respectively. The diesel degradation efficiency was higher in the individual additions of glucose or acetate than in the mixture of glucose and acetate. When P concentration increased from 20 mg/L to 30 mg/L, the diesel degradation efficiency was increased by 7% from 65% to 72%, whereas when P concentration was increased from 30 mg/L to 40 mg/L, there was no increase in diesel degradation. One of the five proteins identified by proteome analysis in the 0.5% diesel-treated samples may be involved in alkane degradation and is known as the cytochrome P450 system. Also, two of the sixteen proteins identified in the 1.5% diesel-treated samples may be implicated in the fatty acid transport system and alcohol dehydrogenation.

Proteomic analysis for the effects of non-saponin fraction with rich polysaccharide from Korean Red Ginseng on Alzheimer's disease in a mouse model

  • Sujin Kim;Yunkwon Nam;Min-jeong Kim;Seung-hyun Kwon;Junhyeok Jeon;Soo Jung Shin;Soyoon Park;Sungjae Chang;Hyun Uk Kim;Yong Yook Lee;Hak Su Kim;Minho Moon
    • Journal of Ginseng Research
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    • v.47 no.2
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    • pp.302-310
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    • 2023
  • Background: The most common type of dementia, Alzheimer's disease (AD), is marked by the formation of extracellular amyloid beta (Aβ) plaques. The impairments of axons and synapses appear in the process of Aβ plaques formation, and this damage could cause neurodegeneration. We previously reported that non-saponin fraction with rich polysaccharide (NFP) from Korean Red Ginseng (KRG) showed neuroprotective effects in AD. However, precise molecular mechanism of the therapeutic effects of NFP from KRG in AD still remains elusive. Methods: To investigate the therapeutic mechanisms of NFP from KRG on AD, we conducted proteomic analysis for frontal cortex from vehicle-treated wild-type, vehicle-treated 5XFAD mice, and NFP-treated 5XFAD mice by using nano-LC-ESI-MS/MS. Metabolic network analysis was additionally performed as the effects of NFP appeared to be associated with metabolism according to the proteome analysis. Results: Starting from 5,470 proteins, 2,636 proteins were selected for hierarchical clustering analysis, and finally 111 proteins were further selected for protein-protein interaction network analysis. A series of these analyses revealed that proteins associated with synapse and mitochondria might be linked to the therapeutic mechanism of NFP. Subsequent metabolic network analysis via genome-scale metabolic models that represent the three mouse groups showed that there were significant changes in metabolic fluxes of mitochondrial carnitine shuttle pathway and mitochondrial beta-oxidation of polyunsaturated fatty acids. Conclusion: Our results suggested that the therapeutic effects of NFP on AD were associated with synaptic- and mitochondrial-related pathways, and they provided targets for further rigorous studies on precise understanding of the molecular mechanism of NFP.

Mining the Proteome of Fusobacterium nucleatum subsp. nucleatum ATCC 25586 for Potential Therapeutics Discovery: An In Silico Approach

  • Habib, Abdul Musaweer;Islam, Md. Saiful;Sohel, Md.;Mazumder, Md. Habibul Hasan;Sikder, Mohd. Omar Faruk;Shahik, Shah Md.
    • Genomics & Informatics
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    • v.14 no.4
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    • pp.255-264
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    • 2016
  • The plethora of genome sequence information of bacteria in recent times has ushered in many novel strategies for antibacterial drug discovery and facilitated medical science to take up the challenge of the increasing resistance of pathogenic bacteria to current antibiotics. In this study, we adopted subtractive genomics approach to analyze the whole genome sequence of the Fusobacterium nucleatum, a human oral pathogen having association with colorectal cancer. Our study divulged 1,499 proteins of F. nucleatum, which have no homolog's in human genome. These proteins were subjected to screening further by using the Database of Essential Genes (DEG) that resulted in the identification of 32 vitally important proteins for the bacterium. Subsequent analysis of the identified pivotal proteins, using the Kyoto Encyclopedia of Genes and Genomes (KEGG) Automated Annotation Server (KAAS) resulted in sorting 3 key enzymes of F. nucleatum that may be good candidates as potential drug targets, since they are unique for the bacterium and absent in humans. In addition, we have demonstrated the three dimensional structure of these three proteins. Finally, determination of ligand binding sites of the 2 key proteins as well as screening for functional inhibitors that best fitted with the ligands sites were conducted to discover effective novel therapeutic compounds against F. nucleatum.

Expression of AGR-2 in Chicken Oviduct during Laying Period

  • Kim, Nam-Soo;Shen, Yan-Nan;Kim, Tae-Yoon;Byun, Sung-June;Jeon, Ik-Soo;Kim, Sang-Hoon
    • BMB Reports
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    • v.40 no.2
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    • pp.212-217
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    • 2007
  • The chicken oviduct is a dynamic organ that produces secretory proteins such as ovalbumin during the laying period. In this study, we identified oviduct-specific proteins in hens during the egg-laying period by proteomic analysis. Proteins extracted from the magnum of hens of different ages (5, 35, and 65 weeks) were analyzed by two-dimensional gel electrophoresis to compare the intensity of proteins among samples. Approximately 300 spots were detected on each gel. Based on the comparison of image gels, we found that the intensity of eight spots in 35-week magnums was increased at least by 2-fold compared with the others. Five of the eight spots were identified as calumenin, acidic ribosomal phosphoproteins (ARP), prohibitin, heart fatty acid-binding protein, and anterior gradient-2 (AGR-2). In particular, ARP and AGR-2 were highly expressed in 35- week magnums compared with 5- and 65-week magnums. In addition, the level of these proteins was consistent with their RNA levels. Expression of AGR-2 mRNA was detected in the mature magnum, whereas no signal was observed in premature tissue. Among various tissues, expression of AGR-2 mRNA was highest in the magnum, high in the isthmus, and five fold lower in muscle. It was undetectable in the liver and in other tissues (heart and kidney). However, the mRNA levels of other proteins were ubiquitous among tissues. In transcriptional activity of AGR-2, a 3.0 kb fragment of promoter region containing potential estrogen receptor binding sites had enhanced its activity strongly. In conclusion, these results suggest that AGR-2 has functional regulatory roles in the chicken oviduct during the egglaying period.

Transcript accumulation of carotenoid biosynthesis genes in the cyanobacterium Synechocystis sp. PCC 6803 during the dark-to-light transition is mediated by photosynthetic electron transport

  • Ryu, Jee-Youn;Song, Ji-Young;Chung, Young-Ho;Park, Young-Mok;Chow, Wah-Soon;Park, Youn-Il
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.149-155
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    • 2010
  • Expression of the genes for carotenoid bio-synthesis (crt) is dependent on light, but little is known about the underlying mechanism of light sensing and signalling in the cyanobacterium Synechocystis sp. PCC 6803 (hereafter, Synechocystis). In the present study, we investigated the light-induced increase in the transcript levels of Synechocystis crt genes, including phytoene synthase (crtB), phytoene desaturase (crtP), ${\zeta}$-carotene desaturase (crtQ), and ${\beta}$-carotene hydroxylase (crtR), during a darkto-light transition period. During the dark-to-light shift, the increase in the crt transcript levels was not affected by mutations in cyanobacterial photoreceptors, such as phytochromes (cph1, cph2 and cph3) and a cryptochrome-type photoreceptor (ccry), or respiratory electron transport components NDH and Cyd/CtaI. However, treatment with photosynthetic electron transport inhibitors significantly diminished the accumulation of crt gene transcripts. Therefore, the light induction of the Synechocystis crt gene expression is most likely mediated by photosynthetic electron transport rather than by cyanobacterial photoreceptors during the dark-to-light transition.