• Title/Summary/Keyword: Subcellular

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Computational Identification of Essential Enzymes as Potential Drug Targets in Shigella flexneri Pathogenesis Using Metabolic Pathway Analysis and Epitope Mapping

  • Narad, Priyanka;Himanshu, Himanshu;Bansal, Hina
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.621-629
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    • 2021
  • Shigella flexneri is a facultative intracellular pathogen that causes bacillary dysentery in humans. Infection with S. flexneri can result in more than a million deaths yearly and most of the victims are children in developing countries. Therefore, identifying novel and unique drug targets against this pathogen is instrumental to overcome the problem of drug resistance to the antibiotics given to patients as the current therapy. In this study, a comparative analysis of the metabolic pathways of the host and pathogen was performed to identify this pathogen's essential enzymes for the survival and propose potential drug targets. First, we extracted the metabolic pathways of the host, Homo sapiens, and pathogen, S. flexneri, from the KEGG database. Next, we manually compared the pathways to categorize those that were exclusive to the pathogen. Further, all enzymes for the 26 unique pathways were extracted and submitted to the Geptop tool to identify essential enzymes for further screening in determining the feasibility of the therapeutic targets that were predicted and analyzed using PPI network analysis, subcellular localization, druggability testing, gene ontology and epitope mapping. Using these various criteria, we narrowed it down to prioritize 5 novel drug targets against S. flexneri and one vaccine drug targets against all strains of Shigella. Hence, we suggest the identified enzymes as the best putative drug targets for the effective treatment of S. flexneri.

Study on Progesterone $6{\alpha}-Steroid$ Hydroxylase from New-born Rat Liver (신생 쥐 간의 Progesterone $6{\alpha}-Steroid$ Hydroxylase에 대한 연구)

  • Jo, Do-Hyun;Park, Yun-Hee;Ryu, Yeon-Woo
    • Applied Biological Chemistry
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    • v.27 no.2
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    • pp.100-106
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    • 1984
  • Five subcellular fractions were obtained by successive centrifugation from the liver of rats within 6 hours of life and characterized by comparing marker compound or marker enzyms. After incubating $3{\beta}$-hydroxy-$5{\alpha}$-pregnan-20-one with the each fraction, the steroids were analyzed by TLC, GLC and GC-MS. A $6{\alpha}$-hydroxylase which hydroxylizes the tetra-hydrogenated compound of progesterone, $3{\beta}$-hydroxy-$5{\alpha}$-pregnan-20-one, was localized in the crude plasma membrane fraction, but not in the microsome fraction. The maximum 6α-hydroxylation was observed at pH 7.0. While this 6α-steroid hydroxylase was not able to hydroxlyze the progesterone, the $3{\alpha}$-isomer was hydroxylized at the $6{\alpha}$-position.

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Behaviour of the soil residues of the bipyridylium herbicide, [$^{14}C$]paraquat in the micro-ecosystem (Micro-ecosystem중 bipyridylium 제초제 paraquat 토양잔류물의 행적)

  • Kwon, Jeong-Wook;Lee, Jae-Koo
    • The Korean Journal of Pesticide Science
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    • v.3 no.1
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    • pp.66-77
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    • 1999
  • In order to elucidate the fate of the residues of the bipyridylium herbicide paraquat in soil, maize plants were grown for 4 weeks on the specially-made pots filled with two different types of soils containing fresh and 6-week-aged residues of [$^{14}C$]paraquat, respectively. The mineralization of [$^{14}C$]paraquat to $^{14}CO_{2}$ during the aging period and the cultivation period of maize plants amounted to $0.13{\sim}0.18%$ and $0.02{\sim}0.17%$, respectively, of the original $^{14}C$ activities. At harvest the roots and shoots contained less than 0.1% and 0.01% of the originally applied $^{14}C$ activities, respectively, whereas the $^{14}C$ activities remaining in soil were more than 97% in both soils. The water extractability of the soil where maize plants were grown for 4 weeks was less than 1.2% of the original $^{14}C$ activities. Most of the non-extractable soil-bound residues of [$^{14}C$]paraquat were incorporated into the humin fraction. Soil pHs during the aging of soil B and after cultivation in all treatments increased. The distribution of the $^{14}C$ activities in subcellular particles of the maize plant roots was the highest in the residue fraction(incompletely homogenized tissue). Dehydrogenase activities increased after vegetation, regardless of soil aging.

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Temporal and subcellular distributions of Cy5.5-labeled hyaluronic acid nanoparticles in mouse organs during 28 days as a drug carrier

  • Lin, Chunmei;Kim, Saet Byeol;Yon, Jung-Min;Park, Seul Gi;Gwon, Lee Wha;Lee, Jong-Geol;Baek, In-Jeoung;Lee, Beom Jun;Yun, Young Won;Nam, Sang-Yoon
    • Korean Journal of Veterinary Research
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    • v.57 no.4
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    • pp.215-222
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    • 2017
  • Temporal and subcellular distributions of hyaluronic acid (HA) as a degradable nanoparticle (NP) in animals were investigated to determine if HA-NP could be utilized as an appropriate drug delivery system. After mice were intravenously injected with 5 mg/kg of Cy5.5-labeled HA-NP sized 350-400 nm or larger HA-polymers, the fluorescence intensity was measured in all homogenized organs from 0.5 h to 28 days. HA-NP was greatly detected in spleen, liver and kidney until day 28, while it was maintained at low levels in other organs. HA-polymer was observed at low levels in all organs. HA-NP quantities in spleen and liver were reduced until day 3, but increased sharply between days 3 and 7, then decreased again, while their HA-polymers were maintained at low levels until day 28. In kidneys, both HA-NP and HA-polymer showed high levels after 0.5 h of administration, but steadily decreased until day 28. According to ultrastructural analyses, HA-NP was engulfed in Kupffer cells of liver and macrophages of spleen and kidney at day 1 and was accumulated in the cytoplasm of kidney tubular cells at day 7. Overall, these findings suggest that HA-NP could be considered a desirable drug carrier in the liver, kidney, or spleen.

Effects of brazilin and haematoxylin on the lipidperoxidation in the rat liver mitochondria

  • Moon, Chang-Kiu;Ha, Bae-Jin;Lee, Chong-Chul
    • Archives of Pharmacal Research
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    • v.7 no.1
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    • pp.63-64
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    • 1984
  • This major sites of liquidperoxidation-damage within the cell are at biomembrances, especially those of subcellular organells such as mitochondria and microsomes whose membranes contain relatively large amount of polyunsaturated fatty acids. Mitochondria are the power plants of eukaryotic cells. Hence their damage by liquid peroxidation can profoundly affect cellular function.

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Prefractionation and Enrichment for the Analysis of Low Aboundance Proteome (극미세 단백질 분석을 위한 프로테옴 분획 농축 기술)

  • 지재웅;변상요
    • KSBB Journal
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    • v.16 no.5
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    • pp.435-441
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    • 2001
  • In spite of the powerfulness for the simultaneous study of proteome expression and post-translational modification, 2-D PAGE has inevitable limitation on detect low aboundant proteins. Since many of the low abundant proteins are likely to have very important regulatiory functions in cells, separation and analysis of low copy number proteins is an important issue in proteome studies and challenge for 2-D techniques. Among various methods developed to detect low abundant proteins, electrophoretic protein prefractionation, chromatographic protein prefractionation, and subcellular fractionation are explained in this paper. Their practical strengths and weaknesses are also explained with current research trends.

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Mass Spectrometry-Based Proteomic Profiling of Pseudopodia of Metastatic Cancer Cells

  • Choi, Sunkyu
    • Mass Spectrometry Letters
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    • v.11 no.2
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    • pp.25-29
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    • 2020
  • Pseudopodia are dynamic actin cytoskeleton-based membrane protrusions of cells that enable directional cell migration. Pseudopodia of cancer cells play key roles in cancer metastasis. Recent studies using pseudopodial subcellular fractionation methodologies combined with mass spectrometry-based proteomic profiling have provided insight into the pseudopodiome that control the protrusions of invasive metastatic cancer cells. This review highlights how to characterize the protein composition of pseudopodia and develop strategies to identify biomarkers or drug candidates that target reduction or prevention of metastatic cancer.

Sequence analysis of LSU rDNA of Alexandrium tamarense/catenella complex from Korean coastal waters

  • Kim, Keunyong;Kim, Chang-Hoon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.252-254
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    • 2001
  • A great deal of effort has been put into the identification of Alexandrium tamarense/fundyense/catenella complex by understanding correlation between morphological and subcellular characteristics. To date, the most promising tool for the study of these species is sequence analyses of rRNA genes that have been useful for various organisms' taxonomy and phylogeny, and its application such as in situ hybridization. (omitted)

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Mitochondrially Targeted Bcl-2 and Bcl-XL Chimeras Elicit Different Apoptotic Responses

  • Liu, Sen;Pereira, Natasha Ann;Teo, Joong Jiat;Miller, Peter;Shah, Priya;Song, Zhiwei
    • Molecules and Cells
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    • v.24 no.3
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    • pp.378-387
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    • 2007
  • The Bcl-2 family of proteins interacts at the mitochondria to regulate apoptosis. However, the anti-apoptotic Bcl-2 and $Bcl-X_L$ are not completely localized to the mitochondria. In an attempt to generate Bcl-2 and $Bcl-X_L$ chimeras that are constitutively localized to the mitochondria, we substituted their C-terminal transmembrane tail or both the C-terminal transmembrane tail and the adjacent loop with the equivalent regions from Bak or Bax mutant (BaxS184V) as these regions determine the mitochondrial localization of Bak and Bax. The effects of these substitutions on subcellular localization and their activities were assessed following expression in HeLa and CHO K1 cells. The substitution of the C-terminal tail or the C-terminal tail and the adjacent loop of Bcl-2 with the equivalent regions from Bak or the Bax mutant resulted in its association with the mitochondria. This change in subcellular localization of Bcl-2 chimeras triggered cells to undergo apoptotic-like cell death. The localization of this Bcl-2 chimera to the mitochondria may be associated with the disruption of mitochondrial membrane potential. Unlike Bcl-2, the loop structure adjacent to the C-terminal tail in $Bcl-X_L$ is crucial for its localization. To localize the $Bcl-X_L$ chimeras to the mitochondria, the loop structure next to the C-terminal tail in $Bcl-X_L$ protein must remain intact and cannot be substituted by the loop from Bax or Bak. The chimeric $Bcl-X_L$ with both its C-terminal tail and the loop structure replaced by the equivalent regions of Bak or Bax mutant localized throughout the entire cytosol. The $Bcl-X_L$ chimeras that are targeted to the mitochondria and the wild type $Bcl-X_L$ provided same protection against cell death under several death inducing conditions.

Cloning and Expression Analysis of a Novel Mouse Zinc Finger Protein Gene Znf313 Abundantly Expressed in Testis

  • Li, Na;Sun, Huaqin;Wu, Qiaqing;Tao, Dachang;Zhang, Sizhong;Ma, Yongxin
    • BMB Reports
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    • v.40 no.2
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    • pp.270-276
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    • 2007
  • We have cloned a novel mouse zinc finger protein gene Znf313 by rapid amplification of cDNA ends (RACE) according to the homologue of human ZNF313 gene. The cDNA is 2,163 base pairs (bp) in length and encodes a 229 amino acids (aa) protein with a $C_3HC_4$ ring finger domain and three $C_2H_2$ domains. 89% and 93% nucleotide (nt) and aa sequence identity is observed with its human homologue. Revealed by Northern blot and RT-PCR, full mRNA consists of 2.16 kb and widely expresses in tissues as a single transcript, most abundantly in heart, liver, kidney and testis. The expression of Znf313 in testis is detected in all development stages. Western blot analysis also reveals that Znf313 is expressed in the tissues. Immunohistochemical staining and subcellular localization demonstrate that Znf313 is expressed both in the cytoplasm and nucleus whereas predominantly localized in the nucleus. Present data suggests that Znf313 gene might play a fundamental role in gene transcription and regulation in organism and relates to spermatogenesis.