• Journal of Korean Neurosurgical Society
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• 제41권5호
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• pp.306-313
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• 2007

Objective : In permanent distal middle cerebral artery occlusion [pdMCAO] model of rats, the temporal order of subcellular translocation is not fully understood yet. We studied translocation sequence of cytochrome c and apoptosis inducing factor [AIF] after pdMCAO and patterns of expression. Methods : Twenty-one male rats - with ten minutes, 1, 4, 8, 24 and 48 hours of pdMCAO groups - were enrolled. At core and penumbra area of each cerebral cortex, Western blotting of cytochrome c and AIF were performed using cytosolic fractions and then compared with sham specimens. With 48 hours group, the expression of cytochrome c and AIF was examined with immunofluorescent staining. Results : Compared to sham, the cytosolic translocation of cytochrome c significantly increased at all time points [p<0.05]. As early as 10 min after onset of ischemia, it was increased significantly [p<0.01]. The cytosolic translocation of AIF showed gradual increase with the passage of time and significantly increased 8 hours after [p<0.05]. As late as 24 hours and 48 hours after onset of ischemia, there were increased most significantly [p<0.01]. At penumbra, both proteins failed to show significant increase at all time points. At 48 hours after ischemia, colocalization of cytochrome c and AIF were confirmed. Conclusion : Cytosolic translocation of cytochrome c peaks much earlier than that of AIF in pdMCAO model of rat. Caspase dependent apoptosis activates soon after ischemia and later, it can be reinforced by gradually increasing AIF in ischemic core.

• BMB Reports
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• 제28권3호
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• pp.254-260
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• 1995

Cytochrome oxidase was purified from bovine-heart mitochondria and its enzymatic properties were examined. The purified cytochrome oxidase was identified by its absorption spectrum and chromatogram through gel filtration. The specific activity, purification degree and yield of purified cytochrome oxidase were 18 nmol/mg/ml/min, 24.83 fold and 0.93%, respectively. The activity of the enzyme assayed by a ferrocytochrome $c-O_2$ system was optimized at $25^{\circ}C$ and pH 6.5. Examining the effect of nonionic detergents established that cytochrome oxidase was deactivated by Triton X-100. The oxidase was activated by Tween 80 and deactivated by Tween 20. The Michaelis constant and maximum velocity of the oxidase for ferrocytochrome c were 0.032~0.044 mM and 0.019~0.021 mM/min, respectively. After adaption to basal diet for a week, experimental diets containing 6 mg Cu/kg, or zero mg Cu/kg, or 12 mg Cu/kg were fed to a control group, a copper-free group and a copper-rich group of Sprague-Dawley rats, respectively, for 4 weeks. The specific activities assayed for the ferrocytochrome $c-O_2$ system of isolated cytochrome oxidase from the rat liver of control, copper-free, and copper-rich group were 1.00, 1.19, and 0.878 nmol/mg/ml/min, respectively. Their degrees of purification were 11.38, 10.82 and 8.78 fold, respectively. The specific activities for liver and heart mitochondrial cytochrome oxidase of copper-free/copper-rich groups assayed using the ferrocytochrome $c-O_2$ system were 81.4% and 96.4%/64.1% and 61.1%, respectively, compared with those of the control.

• 한국자기공명학회논문지
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• 제3권1호
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• pp.12-26
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• 1999

The 1H NMR signals of the heme methyl, propionate and related chemical groups of cytochrome C3 from Desulfovibrio vulgaris Miyazaki F (D.v. MF) were assigned by means of 1D NOE, 2D DQFCOSY and 2D TOCSY spectra. They were consistent with the assignments of the hemes with the highest and second-lowest redox potentials reported by Gayda et al. [Reference: 15]. The heme assignments were also supported by NOE between the methyl groups of these hemes and the side chain of Val-18, All the results contradicted the heme assignments for D.v. MF cytochrome C3 made on the basis of NMR [Reference: 11]. Based on these assignments, the interaction of cytochrome C3 with ferredoxin I was investigated by NMR. The major interaction site of cytochrome C3 was identified as the heme with the highest redox potential, which is surrounded by the highest density of positive charges. The stoichiometry and association constant were two cytochrome C3 molecules per monomer of ferredoxin I and 108 M-2 (at 53 mM ionic strength and $25^{\circ}C$), respectively.

• Bulletin of the Korean Chemical Society
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• 제27권11호
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• pp.1891-1893
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• 2006

• Applied Biological Chemistry
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• 제38권2호
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• pp.168-173
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• 1995

Endosulfan이 흰쥐(Balb/c.) 체내의 cytochrome P-450 효소계에 미치는 영향을 조사하기 위해 endosulfan을 7.5 mg/kg 수준으로 복강주사하였다. Endosulfan을 복강주사 처리하여 48시간 후 적출한 흰쥐의 간에서는 cytochrome P-450 함량이 $3.3{\sim}4.2$배, cytochrome $b_5$ 함량이 $2.3{\sim}3.8$배, NADPH cytochrome P-450 reductase 활성이 $5.3{\sim}6.4$배 그리고 총 haem 함량이 $3.1{\sim}3.6$배씩 대조구의 그것들에 비해 증가하였다. Endosulfan은 대조구 흰쥐 간의 cytochrome P-450 효소계와 상호작용하여 파장 387와 389 nm에서 흡광도의 증가를 보였으며 파장 407 nm에서 넓은 흡수대를 형성하였다. 환원형 P-450-CO spectrum은 대조구의 경우 파장 451 nm에서 흡광도의 극대를 보인 반면 endosulfan으로 처리된 흰쥐 간의 그것은 파장 449와 450 nm에서 흡광도의 극대를 보였다. Endosulfan 처리로 흰쥐 간과 신장의 aldrin epoxidase 활성이 각각 2.8배와 2.1배씩 증가하였으며 7-ethoxyresorufin dealkylase 활성은 각각 1.7배와 1.8배씩 증가하였다.

• Bulletin of the Korean Chemical Society
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• 제26권2호
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• pp.278-280
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• 2005

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 medium containing a small amount of $NH_4C$l 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 $^{15}NH_4$Cl; it was confirmed by $^1H-^{15}N$ HMQC.

• 생약학회지
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• 제20권3호
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• pp.154-161
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• 1989

Effects of panaxydol, panaxynol and panaxytriol isolated from Panax ginseng C.A. Meyer on some enzyme activities were determined. Activities of ATPase, membrane-bound enzyme from Sarcoma 180 and rat liver were slightly inhibited by panaxydol. Activities of 5'-nucleotidase, membrane-bound enzyme and succinate cytochrome c reductase in mitochonidria from sarcoma 180 and rat livers were significantly inhibited in a dose-dependent manner by panaxynol. The inhibitory effects of panaxydol and panaxynol on succinate cytochrome c reductase activities were more potent than those on 5'-nucleotidase activities and panaxynol was found to be a very potent inhibitor of succinate cytochrome c reductase. Activities of glucose-6-phosphatase in endoplasmic reticulum from Sarcoma 180 and rat livers were not affected by all three polyacetylenes. These results suggested that the inhibitory effects of panaxydol and panaxynol on enzyme activities might contribute to their biological activities.

• Animal Systematics, Evolution and Diversity
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• 제28권3호
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• pp.215-219
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• 2012

To examine genetic divergences of two endemic Sorex caecutiens subspecies from Korea (S. c. hallamontanus in Korean Jeju Island and S. c. annexus in the mainland Korean Peninsula), we obtained partial cytochrome oxidase I (COI) sequences (429 bp) and complete cytochrome b sequences (1,140 bp) from the two Korean subspecies, and we compared these sequences to the corresponding sequences of S. caecutiens, obtained from GenBank. We found that Jeju S. c. hallamontanus is one of three clades within S. caecutiens, with an average Jukes-Cantor distance of 1.57% in the COI sequences and the distance of 2.07% and 11 fixed site differences in the cytochrome b sequences, indicating that Jeju S. c. hallamontanus is one endemic subspecies with concordant genetic distinctness, although further analyses with nuclear DNA sequences are necessary to confirm these findings. However, S. c. annexus from the mainland Korean Peninsula was not divergent from S. c. macropygmaeus from northeastern China and adjacent Russia, indicating that S. c. annexus from the mainland Korean Peninsula is another endemic subspecies with only morphological differences, although it is necessary to reexamine the subspecies status of S. c. annexus.

• Animal cells and systems
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• 제13권2호
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• pp.105-112
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• 2009

In this study, the effects of iron on cytochrome c oxidase (CcO) in rat lung mitochondria were examined. Similar to liver mitochondria, iron accumulated considerably in lung mitochondria (more than 2-fold). Likewise, the reactive oxygen species and nitric oxide (NO) content of mitochondria were increased by more than 50% and 100%, respectively. NO might be produced by nitric oxide synthase (NOS), eNOS and iNOS type, with particular contribution by NOS in mitochondria. The respiratory control ratio of iron overloaded lung mitochondria dropped to nearly 50% due to increased state 4. Likewise, cytochrome c oxidase activity was lowered significantly to approximately 50% due to excess iron. Real-time PCR revealed that the expression of isoforms 1 and 2 of subunit IV of CeO was enhanced greatly under excess iron conditions. Taken together, these results show that oxidative phosphorylation within lung mitochondria may be influenced by iron overload through changes in cytochrome c oxidase and NO.

• BMB Reports
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• 제37권4호
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• pp.445-453
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• 2004

We identified apoptosis as being a significant mechanism of toxicity following the exposure of HeLa cell cultures to abrin holotoxin, which is in addition to its inhibition of protein biosynthesis by N-glycosidase activity. The treatment of HeLa cell cultures with abrin resulted in apoptotic cell death, as characterized by morphological and biochemical changes, i.e., cell shrinkage, internucleosomal DNA fragmentation, the occurrence of hypodiploid DNA, chromatin condensation, nuclear breakdown, DNA single strand breaks by TUNEL assay, and phosphatidylserine (PS) externalization. This apoptotic cell death was accompanied by caspase-9 and caspase-3 activation, as indicated by the cleavage of caspase substrates, which was preceded by mitochondrial cytochrome c release. The broad-spectrum caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD-fmk), prevented abrin-triggered caspase activation and partially abolished apoptotic cell death, but did not affect mitochondrial cytochrome c release. These results suggest that the release of mitochondrial cytochrome c, and the sequential caspase-9 and caspase-3 activations are important events in the signal transduction pathway of abrin-induced apoptotic cell death in the HeLa cell line.