Surface functionalization dependent subcellular localization of Superparamagnetic nanoparticle in plasma membrane and endosome

Thimiri Govinda Raj Deepak B.,Khan Niamat Ali 나노기술연구협의회 2018 Nano Convergence Vol.5 No.4

In this article, we elaborate the application of thermal decomposition based synthesis of Fe3O4 superparamagnetic nanoparticle (SPMNP) in subcellular fractionation context. Here, we performed surface functionalization of SPMNP with phospholipids and dimercaptosuccinic acid. Surprisingly, we observed surface functionalization dependent SPMNP localization in subcellular compartments such as plasma membrane, endosomes and lysosomes. By using SPMNP based subcellular localization with pulse–chase methodology, we could use SPMNP for high pure-high yield organelle (plasma membrane, endosomes and lysosome) fractionation. Further, SPMNP that are distinctly localized in subcellular compartments can be used as technology for subcellular fractionation that can complement existing tools for cell biology research. As a future perspective, isolated magnetic organelles can be extended to protein/protein complex purification for biochemical and structural biology studies.

Conversion of Apricot Cyanogenic Glycosides to Thiocyanate by Liver and Colon Enzymes

Jiyeon Lee,Hoonjeong Kwon 한국독성학회 2009 Toxicological Research Vol.25 No.1

Some of the edible plants like apricot kernel, flaxseed, and cassava generate hydrogen cyanide (HCN) when cyanogenic glycosides are hydrolyzed. Rhodanese (thiosulfate: cyanide sulfurtransferases of TSTs; EC: 2.8.1.1) is a sulfide-detoxifying enzymes that converts cyanides into thiocyanate and sulfite. This enzyme exists in a liver and kidneys in abundance. The present study is to evaluate the conversion of apricot cyanogenic glycosides into thiocyanate by human hepatic (HepG2) and colonal (HT-29) cells, and the induction of the enzymes in the rat. The effects of short term exposure of amygdalin to rats have also been investigated. Cytosolic, mitochondrial, and microsomal fractions from HepG2 and HT-29 cells and normal male Spraque-Dawley rats were used. When apricot kernel extract was used as substrate, the rhodanese activity in liver cells was higher than the activity in colon cells, both from established human cell line or animal tissue. The cytosolic fractions showed the highest rhodanese activity in all of the cells, exhibiting two to three times that of microsomal fractions. Moreover, the cell homogenates could metabolize apricot extract to thiocyanate suggesting cellular hydrolysis of cyanogenic glycoside to cyanide ion, followed by a sulfur transfer to thiocyanate. After the consumption of amygdalin for 14 days, growth of rats began to decrease relative to that of the control group though a significant change in thyroid has not been observed. The resulting data support the conversion to thiocyanate, which relate to the thyroid dysfunction caused by the chronic dietary intake of cyanide. Because Korean eats a lot of Brassicaceae vegetables such as Chinese cabbage and radish, the results of this study might indicate the involvement of rhodanese in prolonged exposure of cyanogenic glycosides.

Conversion of Apricot Cyanogenic Glycosides to Thiocyanate by Liver and Colon Enzymes

Lee, Ji-Yeon,Kwon, Hoon-Jeong Korean Society of ToxicologyKorea Environmental Mu 2009 Toxicological Research Vol.26 No.1

Some of the edible plants like apricot kernel, flaxseed, and cassava generate hydrogen cyanide (HCN) when cyanogenic glycosides are hydrolyzed. Rhodanese (thiosulfate: cyanide sulfurtransferases of TSTs; EC: 2.8.1.1) is a sulfide-detoxifying enzymes that converts cyanides into thiocyanate and sulfite. This enzyme exists in a liver and kidneys in abundance. The present study is to evaluate the conversion of apricot cyanogenic glycosides into thiocyanate by human hepatic (HepG2) and colonal (HT-29) cells, and the induction of the enzymes in the rat. The effects of short term exposure of amygdalin to rats have also been investigated. Cytosolic, mitochondrial, and microsomal fractions from HepG2 and HT-29 cells and normal male Spraque-Dawley rats were used. When apricot kernel extract was used as substrate, the rhodanese activity in liver cells was higher than the activity in colon cells, both from established human cell line or animal tissue. The cytosolic fractions showed the highest rhodanese activity in all of the cells, exhibiting two to three times that of microsomal fractions. Moreover, the cell homogenates could metabolize apricot extract to thiocyanate suggesting cellular hydrolysis of cyanogenic glycoside to cyanide ion, followed by a sulfur transfer to thiocyanate. After the consumption of amygdalin for 14 days, growth of rats began to decrease relative to that of the control group though a significant change in thyroid has not been observed. The resulting data support the conversion to thiocyanate, which relate to the thyroid dysfunction caused by the chronic dietary intake of cyanide. Because Korean eats a lot of Brassicaceae vegetables such as Chinese cabbage and radish, the results of this study might indicate the involvement of rhodanese in prolonged exposure of cyanogenic glycosides.

Subcellular Localization of Catalase Encoded by the ctl+ Gene in Schizosaccharomyces pombe

Lee, Sang-il,Lee, Joon,Roe, Jung-Hye The Microbiological Society of Korea 2000 The journal of microbiology Vol.38 No.3

The cttl+ gene in Schizosaccharomyces pombe encoeds a catalse responsible for H2O2-resistance of this organism as judged by the H2O2-sensitive phenotype of the ctt1Δ mutant. In this study, we investigated the subcellular localization of the Ctt1 gene product. In wild type cells catalase activity was detected in the organelle fraction as well as in the cytosol. The ctt1Δ mutant contained no catalase activity, indicating that both cytosolic and organellar catalases are the products of a single ctt1+ gene. Western bolt analysis revealed two catalase bands, both of which disappeared in the ctt1Δ mutant. The major, fastermigrating band existed in the cytosol whereas the monor, slower-migrating band appeared to be located in organelles, most likely in peroxisomes. These results suggest that the ctt1+ gene product targeted to the peroxisome is a modified form of the one in the cytosol.

The influence of endogenous cathepsin in different subcellular fractions on the quality deterioration of Northern pike (Esox lucius) fillets during refrigeration and partial freezing storage

Hengheng Qiu,Xin Guo,Xiaorong Deng,Xiaobing Guo,Xiaoying Mao,Chengjian Xu,Jian Zhang 한국식품과학회 2020 Food Science and Biotechnology Vol.29 No.10

The purpose of this study was to investigate theendogenous cathepsin activity in each subcellular fractionand the effect of this activity on myofibrillar protein andtexture during refrigeration and partial freezing storage ofnorthern pike (Esox lucius) fillets. The results showed thatfillets stored under the refrigerated condition were moresusceptible to oxidation than partial freezing. Endogenouscathepsin activity indicated that partial freezing destroysthe integrity of lysosomes more effectively than refrigerationand inhibits the increase in cathepsin B and B ? L inlysosomes. The activity of cathepsin B and B ? L inlysosomes, mitochondria and myofibrils under the partialfreezing conditions was always lower than that underrefrigeration. Texture analysis showed that refrigerationhad a negative impact on hardness and springiness. Inconclusion, the cathepsin activity in each subcellularfraction was effectively inhibited and better textural characteristicswere obtained with partial freezing thanrefrigeration.

Endocytic Rab proteins are required for hepatitis C virus replication complex formation

Manna, D.,Aligo, J.,Xu, C.,Park, W.S.,Koc, H.,Do Heo, W.,Konan, K.V. Academic Press 2010 Virology Vol.398 No.1

During infection, hepatitis C virus (HCV) NS4B protein remodels host membranes to form HCV replication complexes (RC) which appear as foci under fluorescence microscopy (FM). To understand the role of Rab proteins in forming NS4B foci, cells expressing the HCV replicon were examined biochemically and via FM. First, we show that an isolated NS4B-bound subcellular fraction is competent for HCV RNA synthesis. Further, this fraction is differentially enriched in Rab1, 2, 5, 6 and 7. However, when examined via FM, NS4B foci appear to be selectively associated with Rab5 and Rab7 proteins. Additionally, dominant negative (DN) Rab6 expression impairs Rab5 recruitment into NS4B foci. Further, silencing of Rab5 or Rab7 resulted in a significant decrease in HCV genome replication. Finally, expression of DN Rab5 or Rab7 led to a reticular NS4B subcellular distribution, suggesting that endocytic proteins Rab5 and Rab7, but not Rab11, may facilitate NS4B foci formation.

흰쥐 간장에 있어서 아실-CoA 합성효소4의 기능연구

정영희(Jeong Young Hee),문승주(Moon Seung Ju),강만종(Kang Man Jong) 韓國營養學會 2003 Journal of Nutrition and Health Vol.36 No.4

DNA Repair Synthesis and Ligation Affect the Processing of Excised Oligonucleotides Generated by Human Nucleotide Excision Repair

Kemp, Michael G.,Gaddameedhi, Shobhan,Choi, Jun-Hyuk,Hu, Jinchuan,Sancar, Aziz American Society for Biochemistry and Molecular Bi 2014 The Journal of biological chemistry Vol.289 No.38

<P>Ultraviolet (UV) photoproducts are removed from genomic DNA by dual incisions in humans in the form of 24- to 32-nucleotide-long oligomers (canonical 30-mers) by the nucleotide excision repair system. How the small, excised, damage-containing DNA oligonucleotides (sedDNAs) are processed in cells following the dual incision event is not known. Here, we demonstrate that sedDNAs are localized to the nucleus in two biochemically distinct forms, which include chromatin-associated, transcription factor II H-bound complexes and more readily solubilized, RPA-bound complexes. Because the nuclear mobility and repair functions of transcription factor II H and RPA are influenced by post-incision gap-filling events, we examined how DNA repair synthesis and DNA ligation affect sedDNA processing. We found that although these gap filling activities are not essential for the dual incision/sedDNA generation event <I>per se</I>, the inhibition of DNA repair synthesis and ligation is associated with a decrease in UV photoproduct removal rate and an accumulation of RPA-sedDNA complexes in the cell. These findings indicate that sedDNA processing and association with repair proteins following the dual incisions may be tightly coordinated with gap filling during nucleotide excision repair <I>in vivo</I>.</P>