• Title/Summary/Keyword: Cytochrome C

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Interaction of Cytochrome c and $Mn^{2+}$ -Cytochrome c Peroxidase

  • Kim, Mun-kyoung;M. Kwon;Kim, K.;Sanghwa Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 1999.06a
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    • pp.44-44
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    • 1999
  • Yeast cytochrome c peroxidase (CcP) was cloned and overexpressed in E. coli, and purified by a Ni$^{2+}$-affinity column. HoloCcP was obtained by reconstituting apoCcP with Mn$^{3+}$-protoporphyrin IX (MnPP). Electron paramagnetic resonance (EPR) spectra of spin-labeled holoCcP showed a slightly more immobilized signal than spin-labeled apoCcP.(omitted)

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Expression and Purification of GFPuv/Cytochrome c-552 Fusion Protein in E. coli

  • Hong, Eul-Jae;Lee, Sang-On;Choe, Jeong-U;Hong, Eok-Gi
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.550-553
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    • 2003
  • The genes of GFPuv and Cytochrome c-552 were amplified by using PCR, and then, fused each other. Fusion gene of GFPuv and Cytochrome c-552 was inserted into the pTrcHis B vector and transferred to E. coli. A fusion protein of GFPuv and Cytochrome c-552 was expressed in JM109 and BL21. This fusion protein was composed of a His-tag for the rapid one-step purification using an immobilized metal affinity chromatography.

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Local Photoswitching Effects of Cytochrome c/Viologen/GFP Hetero-Thin Film

  • Yu, Chang-Jun;Choe, Jeong-U;Park, Se-Jeong;Nam, Yun-Seok;O, Byeong-Geun;Lee, Won-Hong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.823-826
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    • 2001
  • In the initial process of photosynthesis, a biological electron transfer system, photoelectric conversion occurs and then long-range electron transfer takes place very efficiently in one direction through the biomolecules. The metal/insulator/metal structured device consisting of GFP, viologen, cytochrome c hetero-thin film was presented based on the biomimesis. GFP, viologen, and cytochrome c was used as an electron sensitizer, a mediator, and an electron acceptor. Cytochrome c molecules and viologen molecules were deposited by Langmuir-Blodgett (LB) technique, and GFP molecules were adsorbed by self-assembly method (SAM). Surface morphology of hetero-thin film was analyzed by scanning tunneling microscopy (STM). Local photoswitching effects of a proposed photodiode were verified by current-voltage measurements using hybrid STM/I-V measurement system.

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Effects of pH on Purification of GFPuv/Cytochrome c-552 Fusion Protein

  • Lee, Sang-On;Hong, Eul-Jae;Choe, Jeong-U;Hong, Eok-Gi
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.539-542
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    • 2003
  • Fusion gene of GFPuv and Cytochrome c-552 was inserted into the pTrcHis B vector and transferred to E. coli. A fusion protein of GFPuv and Cytochrome c-552 was expressed in BL21. This fusion protein was composed of a His-tag for purification using an immobilized metal affinity chromatography(IMAC). IMAC constitutes a rather facile means of unravelling the principles of recognition and, in particular, of identifying the counterligands on the protein surface, which interact with the ligated and immobilized metal ions. Histidine when present on the surface of a protein molecule under a favorable solvent condition, may serve as electron donors in coordination with the immobilized chelates of some transition metal ions$(Ni^{2+})$.

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Inhibitory Effects of Exogenous Cu2+ and Zn2+ on the Cytochrome c Oxidase Activity

  • Min, Tong-Pil;Han, Sang-Hwa
    • BMB Reports
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    • v.28 no.4
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    • pp.311-315
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    • 1995
  • Exogenous $Cu^{2+}$ or $Zn^{2+}$ at micromolar concentration had a strong inhibitory effect on detergent-solubilized cytochrome c oxidase. A similar effect was observed when $Cu^{2+}$ was added to vesicular cytochrome c oxidase, although the extent of inhibition was significantly larger for the uncoupled state than for the coupled state. Interestingly, the inhibition by $Zn^{2+}$ was almost negligible for both the coupled and uncoupled states. These results suggest that the binding sites for $Cu^{2+}$ ions are exposed to the extravesicular side. whereas those for $Zn^{2+}$ are exposed to the matrix side. The EPR spectra of bound $Cu^{2+}$ ions at 77 K indicate that each of the first two $Cu^{2+}$ ions is ligated by three or four histidine residues, as evidenced by distinct $^{14}N$ superhyperfine splitting. These $Cu^{2+}$ ions can not be removed readily by EDTA and inhibit the enzyme activity by as much as 80%.

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Stable Isotope Labeled Cytochrome $c_3$ from Desulfovibrio vulgaris on a Defined Medium as Sole Nitrogen Source

  • Kim, Andre;Shim, Yoon-Bo;Kang, Shin-Won;Park, Jang-Su
    • BMB Reports
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    • v.33 no.6
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    • pp.506-509
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    • 2000
  • To obtain Cytochrome $c_3$ labeled with a stable isotope, the conditions of cultivation and the composition of medium for DvMF were examined. The growth of DvMF was steady and reproducible under purging with $N_2$ and under pH control. DvMF was able to go on a defined medium without natural products. The composition of the medium containing a small amount of $NH_4Cl$ as sole nitrogen source was established. Then, uniformly $^{15}N-labeled$ Cytochrome $c_3$ was obtained during the culture of DvMF in a defined medium with $^{15}NH_4Cl$; it was confirmed by $^{1}H-^{15}N$ HMQC.

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Protective Effects of Histidine Dipeptides on the Modification of Neurofilament-L by the Cytochrome c/Hydrogen Peroxide System

  • Kim, Nam-Hoon;Kang, Jung-Hoon
    • BMB Reports
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    • v.40 no.1
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    • pp.125-129
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    • 2007
  • Neurofilament-L (NF-L) is a major element of the neuronal cytoskeleton and is essential for neuronal survival. Moreover, abnormalities in NF-L result in neurodegenerative disorders. Carnosine and the related endogeneous histidine dipeptides prevent protein modifications such as oxidation and glycation. In the present study, we investigated whether histidine dipeptides, carnosine, homocarnosine, or anserine protect NF-L against oxidative modification during reaction between cytochrome c and $H_2O_2$. Carnosine, homocarnosine and anserine all prevented cytochrome c/$H_2O_2$-mediated NF-L aggregation. In addition, these compounds also effectively inhibited the formation of dityrosine, and this inhibition was found to be associated with the reduced formations of oxidatively modified proteins. Our results suggest that carnosine and histidine dipeptides have antioxidant effects on brain proteins under pathophysiological conditions leading to degenerative damage, such as, those caused by neurodegenerative disorders.

$^1H$ NMR Estimation of Multi-Redox potentials of Cytochrome $c_3$ from Desulfovibrio vulgaris Hildenborough

  • 박장수;강신원;최성낙
    • Bulletin of the Korean Chemical Society
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    • v.16 no.4
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    • pp.331-336
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    • 1995
  • The macroscopic and microscopic redox potentials of tetrahemoprotein, cytochrome c3 from Desulfovibrio vulgaris(Hildenborough) (DvH) were estimated from 1H NMR and differential pulse polarography(DPP). Five sets of NMR resonances were confirmed by a redox titration. They represent cytochrome c3 molecules in five macroscopic redox states. The electron transfer in cytochrome c3 involves four consecutive one-electron steps. The saturation transfer method was used to determine the chemical shifts of eight heme methyl resonances in five different oxidation states. Thirty two microscopic redox potentials were estimated. The results showed the presence of a strong positive interaction between a pair of particular hemes. Comparing the results with those of Desulfovibrio vulgaris Miyazaki F (DvMF), it was observed that the two proteins resemble each other in overall redox pattern, but there is small difference in the relative redox potentials of four hemes.

Effect of Cypermethrin and Piperonyl Butoxide on Toxic Response in Rats (Cypermethrin과 Piperonyl butoxide가 rat의 독성반응에 미치는 영향)

  • Chung, Kyu-Hyuok;Hong, Sa-Uk
    • YAKHAK HOEJI
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    • v.34 no.2
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    • pp.69-79
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    • 1990
  • The aim of this experiment is to observe the toxicity of cypermethrin[S, R- -cyano-3-phenoxybenzyl-(1R, 1s, cis, trans)-2,2-dimethyl-3-(2,2-dichlorovinyl) cyclopropane carboxylate]and to investigate the synergistic effect of piperonyl butoxide on the cypermethrin toxicity. In cypermethrin (CYP) treated group, the biochemical parameters such as ALT, LDH, glucose in serum were remarkably elevated. The content of cytochrome P-450 and activity of NADPH-cytochrome c reductase in renal microsomal fraction were increased but those in hepatic microsomal fraction were not significantly increased. The activity of aniline hydroxylase and ATPase in liver were decreased. In the case of CYP plus piperonyl butoxide (PB) treated group, AST, ALT, LDH and glucose were more increased. Cytochrome P-450 and NADPH-cytochrome c reductase in liver and kidney were supressed and aniline hydroxylase and ATPase in liver were more decreased. Especially, in the case of CYP plus PB 100 mg/kg treated group, hepatic TBA value was increased but activity of glucose-6-phosphatase was remarkably depressed.

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Cytochrome C Release and Caspase Activation Induced by 3-Deazaadenosisne is Inhibited by Bcl-2

  • Lee Yong-Joon;Choi Mi-Hyun;Lee Jung-Hee;Kim Ho-Shik;Lee Jeong-Hwa
    • Biomedical Science Letters
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    • v.12 no.2
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    • pp.57-63
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    • 2006
  • Deazaadenosine analogs such as 3-deazaadenosine (DZA), 3-deazaaristeromycin (DZAri) and ara-3-deazaadenine (DZAra-A) were developed as inhibitors of S-adenosylhomocysteine (Ado-Hcy) hydrolase (EC 3.3.1.1). These analogs were reported to induce apoptosis in human and murine leukemic cells. But, the mechanism involved in this apoptosis was not clarified yet. In the present study, we analyze the apoptosis induced by deazaadenosine analogs in human cervival cancer cell line, HeLa and the effect of Bcl-2 on this apoptosis. Whereas neither DZAri nor DZAra-A showed inhibitory effect on HeLa cell growth, DZA induced apoptosis in HeLa cells accompanied by cytochrome c release and activation of various caspases such as caspase-2,-8,-9 and -3. In HeLa-bcl-2 cell line, a stable transfectant of HeLa cell to overexpress Bcl-2, cytochrome c release, activation of all these caspases and the resulted apoptosis by DZA were completely prevented. By in vitro assay of cytochrome c release, in addition, DZA induced cytochrome c release from purified mitochondria of HeLa-pcDNA3 cells, but not HeLa-bcl-2 cells, even in the absence of cytosolic fraction. Therefore, it can be suggested that DZA might damage directly mitochondria leading to activate intrinsic pathway of caspase and thus induce apoptosis. DZA-induced apoptosis in HeLa cells may be in a bcl-2-inhibitable manner and irrelative of Ado-Hcy hydrolase.

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