• Title/Summary/Keyword: Subcellular

Search Result 317, Processing Time 0.026 seconds

The subcellular distribution of MnSOD alters during sodium selenite-induced apoptosis

  • Guan, Liying;Jiang, Qian;Li, Zhushi;Huang, Fang;Ren, Yun;Yang, Yang;Xu, Caimin
    • BMB Reports
    • /
    • v.42 no.6
    • /
    • pp.361-366
    • /
    • 2009
  • It was reported that high doses of sodium selenite can induce apoptosis of cancer cells, but the molecular mechanisms are poorly understood. Manganese superoxide dismutase (MnSOD) converts superoxide radical to hydrogen peroxide within the mitochondrial matrix and is one of the most important antioxidant enzymes. In this study, we showed that 20 ${\mu}M$ sodium selenite could alter subcellular distribution of MnSOD, namely a decrease in mitochondria and an increase in cytosol. The alteration of subcellular distribution of MnSOD is dependent on the production of superoxide induced by sodium selenite.

Subcellular Distribution of Heavy Metals in Organs of Bivalve Modiolus Modiolus Living Along a Metal Contamination Gradient

  • Podgurskaya, Olga V.;Kavun, Victor Ya.
    • Ocean Science Journal
    • /
    • v.41 no.1
    • /
    • pp.43-51
    • /
    • 2006
  • Concentration and distribution of Fe, Zn, Cu, Cd, Mn, Pb, Ni among subcellular fractions (cellular membrane structures and cytosol) and Zn, Cu, Cd among cytoplasmic proteins in the kidney and digestive gland of mussel Modiolus modiolus living along a polymetallic concentration gradient were studied. It was found in the kidney of M. modiolus from contaminated sites that the Fe percent increased in the "membrane" fraction, whereas Zn, Pb, Ni and Mn percent increased in the cytosol compared to the kidney of the control mussel. Note kidney cytosol of M. modiolus from clean and contaminated sites sequestered major parts of Cu and Cd. In the digestive gland of M. modiolus from contaminated sites Fe, Zn, Cd, Mn, Ni percent increased in the "membrane" fraction, whereas Cu, Pb percent increased in the cytosol compared to digestive gland of control mussel. Gel-filtration chromatography shows kidney of M. modiolus contains increased metallothionein-like protein levels irrespective of ambient dissolved metal concentrations. It was shown that the metal detoxification system in the kidney and digestive gland of M. modiolus was efficient under extremely high ambient metal levels. However, under complex environmental contamination in the kidney of M. modiolus, the metal detoxification capacity of metallothionein-like proteins was damaged.

Identification of Novel Regulators of Apoptosis Using a High-Throughput Cell-based Screen

  • Park, Kyung Mi;Kang, Eunju;Jeon, Yeo-Jin;Kim, Nayoung;Kim, Nam-Soon;Yoo, Hyang-Sook;Yeom, Young Il;Kim, Soo Jung
    • Molecules and Cells
    • /
    • v.23 no.2
    • /
    • pp.170-174
    • /
    • 2007
  • High-throughput subcellular imaging is a powerful tool for investigating the function of genes. In order to identify novel regulators of apoptosis we transiently transfected HeLa cells with 938 hypothetical genes of unknown function, and captured their nuclear images with an automated fluorescence microscope. We selected genes that induced greater than 3-fold increase in the percentage of apoptotic nuclei compared with vector-transfected cells. The full-length genes C10orf61, MGC 26717, and FLJ13855 were identified as candidate proapoptotic genes, and their apoptotic effects were confirmed by DNA fragmentation ELISAs and Western blotting for caspase-7 and PARP. We conclude that a subcellular image-based apoptotic screen is useful for identifying genes with proapoptotic activity.

Subcellular Characterization of Porcine Oocytes with Different Glucose-6-phosphate Dehydrogenase Activities

  • Fu, Bo;Ren, Liang;Liu, Di;Ma, Jian-Zhang;An, Tie-Zhu;Yang, Xiu-Qin;Ma, Hong;Zhang, Dong-Jie;Guo, Zhen-Hua;Guo, Yun-Yun;Zhu, Meng;Bai, Jing
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.28 no.12
    • /
    • pp.1703-1712
    • /
    • 2015
  • The in vitro maturation (IVM) efficiency of porcine embryos is still low because of poor oocyte quality. Although brilliant cresyl blue positive (BCB+) oocytes with low glucose-6-phosphate dehydrogenase (G6PDH) activity have shown superior quality than BCB negative (-) oocytes with high G6PDH activity, the use of a BCB staining test before IVM is still controversial. This study aimed to shed more light on the subcellular characteristics of porcine oocytes after selection using BCB staining. We assessed germinal vesicle chromatin configuration, cortical granule (CG) migration, mitochondrial distribution, the levels of acetylated lysine 9 of histone H3 (AcH3K9) and nuclear apoptosis features to investigate the correlation between G6PDH activity and these developmentally related features. A pattern of chromatin surrounding the nucleoli was seen in 53.0% of BCB+ oocytes and 77.6% of BCB+ oocytes showed peripherally distributed CGs. After IVM, 48.7% of BCB+ oocytes had a diffused mitochondrial distribution pattern. However, there were no significant differences in the levels of AcH3K9 in the nuclei of blastocysts derived from BCB+ and BCB- oocytes; at the same time, we observed a similar incidence of apoptosis in the BCB+ and control groups. Although this study indicated that G6PDH activity in porcine oocytes was correlated with several subcellular characteristics such as germinal vesicle chromatin configuration, CG migration and mitochondrial distribution, other features such as AcH3K9 level and nuclear apoptotic features were not associated with G6PDH activity and did not validate the BCB staining test. In using this test for selecting porcine oocytes, subcellular characteristics such as the AcH3K9 level and apoptotic nuclear features should also be considered. Adding histone deacetylase inhibitors or apoptosis inhibitors into the culture medium used might improve the efficiency of IVM of BCB+ oocytes.

Biochemical Properties and Localization of the β-Expansin OsEXPB3 in Rice (Oryza sativa L.)

  • Lee, Yi;Choi, Dongsu
    • Molecules and Cells
    • /
    • v.20 no.1
    • /
    • pp.119-126
    • /
    • 2005
  • ${\alpha}$-Expansins are bound to the cell wall of plants and can be solubilized with an extraction buffer containing 1 M NaCl. Localization of ${\alpha}$-expansins in the cell wall was confirmed by immunogold labeling and electron microscopy. The subcellular localization of vegetative ${\beta}$-expansins has not yet been studied. Using antibodies specific for OsEXPB3, a vegetative ${\beta}$-expansin of rice (Oryza sativa L.), we found that OsEXPB3 is tightly bound to the cell wall and, unlike ${\alpha}$-expansins, cannot be solubilized with extraction buffer containing 1 M NaCl. OsEXPB3 protein could only be extracted with buffer containing SDS. The subcellular localization of the OsEXPB3 protein was confirmed by immunogold labeling and electron microscopy. Gold particles were mainly distributed over the primary cell walls. Immunohistochemistry showed that OsEXPB3 is present in all regions of the coleoptile and root tissues tested.

Sequence driven features for prediction of subcellular localization of proteins

  • Kim, Jong-Kyoung;Bang, Sung-Yang;Choi, Seung-Jin
    • Proceedings of the Korean Society for Bioinformatics Conference
    • /
    • 2005.09a
    • /
    • pp.237-242
    • /
    • 2005
  • Predicting the cellular location of an unknown protein gives a valuable information for inferring the possible function of the protein. For more accurate prediction system, we need a good feature extraction method that transforms the raw sequence data into the numerical feature vector, minimizing information loss. In this paper, we propose new methods of extracting underlying features only from the sequence data by computing pairwise sequence alignment scores. In addition, we use composition based features to improve prediction accuracy. To construct an SVM ensemble from separately trained SVM classifiers, we propose specificity based weighted majority voting. The overall prediction accuracy evaluated by the 5-fold cross-validation reached 88.53% for the eukaryotic animal data set. By comparing the prediction accuracy of various feature extraction methods, we could get the biological insight on the location of targeting information. Our numerical experiments confirm that our new feature extraction methods are very useful for predicting subcellular localization of proteins.

  • PDF

Seeds as Repositories of Recombinant Proteins in Molecular Farming

  • Moloney, Maurice M.
    • Korean Journal of Plant Tissue Culture
    • /
    • v.27 no.4
    • /
    • pp.283-297
    • /
    • 2000
  • Seeds are an ideal repository for recombinant proteins in molecular farming applications. However, in order to use plant seeds efficiently for the production of such proteins, it is necessary to understand a number of fundamental biological properties of seeds. This includes a full understanding of promoters which function in a seed-specific manner, the subcellular targeting of the desired polypeptide and the final form in which a protein is stored. Once a biologically active protein has been deposited in a seed, it is also critical that the protein can be extracted and purified efficiently. In this review, these issues are examined critically to provide a number of approaches which may be adopted for production of recombinant proteins in plants. Particular attention is paid to the relationship between subcellular localization and protein extraction and purification. The robustness and flexibility of seed-based production is illustrated by examples close to or already in commercial production.

  • PDF

Studies on the Intracellular Localization of Polyamines and Their related Enzymes in Spinach Leaves (Polyamine과 Polyamine의 생합성에 관련된 효소들의 시금치잎 세포내 분포에 관한 연구)

  • 김성호
    • Journal of Plant Biology
    • /
    • v.32 no.4
    • /
    • pp.285-292
    • /
    • 1989
  • The intracellular localizations of polyamines and their related enzymes were investigated from young spinach leaves. Polyamines were present in all parts of plant cells, both in the subcellular organelles and in the soluble fraction of cytoplasm, however, polyamines were mainly located in the cytosolic fraction. Most activities of L-arginine decarboxylase(ADC) and L-ornithine decarboxylase(ODC), two important enzymes of putrescine and polyamine biosynthesis, were detected in cytosol fraction, while in subcellular organelles the activities were very low. Activities of diamine oxidase(DAO) and polyamine oxidase(PAO), the catabolic enzyme of diamine and polyamine, were not detected in spinach leaves. It was suggested that polyamines and their related synthetic enzymes were located in the soluble fraction of cytoplasm.

  • PDF

Effects of Extrahepatic Cholestasis on Hepatic $\alpha$-D-Mannosidase Activity in Chronic Ethanol Intoxicated Rats

  • Si-Woo Bae;Chun-Sik Kwak;Chong-Guk Yoon
    • Biomedical Science Letters
    • /
    • v.9 no.1
    • /
    • pp.21-27
    • /
    • 2003
  • Hepatic subcellular $\alpha$-D-mannosidases activities and its Km and Vmax values were determined in chronic ethanol intoxicated rats with extrahepatic cholestasis induced by common bile duct ligation to manifest the biochemical background of alcohol drinking hazard under the hepatobiliary disease. In case of extrahepatic cholestasis, chronic ethanol intoxication in animals led to the increased activities of liver Golgi and microsomal $\alpha$-D-mannosidase as well as the Vmax values of these enzymes. However, the difference of Km values on hepatic subcellular enzymes were not found between the experimental groups. Therefore, the results indicate that the liver Golgi and microsomal $\alpha$-D-mannosidase may be more induced in chronic ethanol intoxication animals in case of cholestasis. Accordingly, the resulting data supported the fact that alcoholic drinks may led to enhancement of the hepatobiliary liver damage.

  • PDF