• BMB Reports
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• 제48권6호
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• pp.354-359
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• 2015

Vascular smooth muscle cells (VSMCs) undergo death during atherosclerosis, a widespread cardiovascular disease. Recent studies suggest that oxidative damage occurs in VSMCs and induces atherosclerosis. Here, we analyzed oxidative damage repair in VSMCs and found that VSMCs are hypersensitive to oxidative damage. Further analysis showed that oxidative damage repair in VSMCs is suppressed by a low level of poly (ADP-ribosyl)ation (PARylation), a key post-translational modification in oxidative damage repair. The low level of PARylation is not caused by the lack of PARP-1, the major poly(ADP-ribose) polymerase activated by oxidative damage. Instead, the expression of poly(ADP-ribose) glycohydrolase, PARG, the enzyme hydrolyzing poly(ADP-ribose), is significantly higher in VSMCs than that in the control cells. Using PARG inhibitor to suppress PARG activity facilitates oxidative damage-induced PARylation as well as DNA damage repair. Thus, our study demonstrates a novel molecular mechanism for oxidative damage-induced VSMCs death. This study also identifies the use of PARG inhibitors as a potential treatment for atherosclerosis. [BMB Reports 2015; 48(6): 354-359]

• 한국동물학회지
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• 제39권3호
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• pp.248-256
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• 1996

본 연구의 목적은 인간의 Poly(ADP-ribose) synthetase (PARS)의 cDNA를 클로닝하여 발현시키기 위해 수행하였다. 먼저, 인간의 PARS cDNA 전체를 포함한 pPARS3.1을 pGEM-7Zf(+) 등의 발현 벡터 클로닝하였다. 이 결과로 생성된 재조합 플라스미드 pPARS6.1이 인간의 PARS cDNA 전체를 포함하고, 올바른 방향으로 삽입되었는지를 확인하기 위해 제한효소 지도를 작성하였고, random primed DNA probe을 이용한 Southern blot 분석에 의해서 PARS가 클로닝되었다는 것을 확인하였다. 또한, 염기서열 분석 결과, 단백질 합성이 시작되는 유전 암호가 정확한 순서로 위치하고 있음을 확인하였다. 재조합된 pPARS6.1의 발현을 위해 배양시 0.4 mM IPTG로 처리하여 만들어진 인간의 PARS 단백질이 10% SDS-PAGE에서 120 kDa 위치에 이동하였다는 것을 nick-translated된 DNA를 probe로 이용하여 확인하였고, Southwestern blot 실험 결과 120 kDa과 98kDa에 위치하는 단백질이 DNA와 결합함을 알 수 있었다.

• Journal of Magnetics
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• 제19권1호
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• pp.20-27
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• 2014

This study examined the relationships between protein expression and Poly ADP ribose polymerase in brain cell death in brains damaged by thrombotic stroke and treated with the Full Wave- Cockroft Walton (FWCW) method of Transcranial Magnetic Stimulation (TMS). The two-way switching element for TMS drove a half-bridge inverter of the current resonance of direct current voltage (+) and direct current voltage (-), and the experiment was conducted by stimulating the mice with thrombotic stroke through a range of pulses. Thrombotic stroke was caused of ligation of the common carotid artery of male SD mice, and blood reperfusion was conducted five minutes later. Protein expression was examined in immune reaction cells, which reacted to an antibody to Poly ADP ribose polymerase in the cerebrum cells, and western blotting. Observations of the PARP changes after thrombotic stroke showed that the number of Poly ADP ribose polymerase reactions were significantly lower (p < 0.05) in the group treated with TMS of the FWCW than the group with thrombotic stroke 24 hours after its onset. The application of FWCW-TMS helped prevent the necrosis of nerve cells and might prevent the brain damage that occurs as a result of thrombotic stroke, and improve the function recovery and disorder of brain cells.

• Molecules and Cells
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• 제38권4호
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• pp.312-317
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• 2015

Depletion of intracellular zinc by N,N,N,N-tetrakis(2-pyridylmethyl) ethylenediamine (TPEN) induces p53-mediated protein synthesis-dependent apoptosis of mouse cortical neurons. Here, we examined the requirement for poly(ADP-ribose) polymerase (PARP)-1 as an upstream regulator of p53 in zinc depletion-induced neuronal apoptosis. First, we found that chemical inhibition or genetic deletion of PARP-1 markedly attenuated TPEN-induced apoptosis of cultured mouse cortical neurons. Poly(ADP-ribosyl)ation of p53 occurred starting 1 h after TPEN treatment. Suggesting the critical role of PARP-1, the TPEN-induced increase of stability and activity of p53 as well as poly(ADP-ribosyl)ation of p53 was almost completely blocked by PARP inhibition. Consistent with this, the induction of downstream pro-apoptotic proteins PUMA and NOXA was noticeably reduced by chemical inhibitors or genetic deletion of PARP-1. TPEN-induced cytochrome C release into the cytosol and caspase-3 activation were also blocked by inhibition of PARP-1. Taken together, these findings indicate that PARP-1 is essential for TPEN-induced neuronal apoptosis.

• 한국동물학회지
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• 제26권3호
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• pp.171-179
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• 1983

MMS와 자외선에 의한 DNA의 절제회복과 단사절단에 미치는 poly(ADP-ribose) polymerase의 저해제인 3-aminobenzamide의 영향을 CHO 세포를 재로로 조사하여 다음과 같은 결과를 얻었다. 1. MMS에 의한 비주기성 DNA 합성률과 DNA 단사 절단률은 이 저해제에 의해 모두 증가하였다. 이는 poly (ADP-ribose) polymeraserk MMS에 의해 유발된 염기 절제회복의 incision step를 억제하는 결과라 생각된다. 2. 자외선에 의한 비주기성 DNA 합성률과 DNA단사 절단률은 이 저해제에 의해 모두 감소하였다. 이는 poly(ADP-ribose) polymerase가 자외선에 의해 유발된 nucleotide 절제회복의 incision step을 돕는 작용을 하는 결과로 생각된다. 3. MMS와 자외선을 복합처리한 실험군에서는, DNA 단사 절단률은 이 저해제에 의해 영향을 받지 않았으며, 비주기성 DNA 합성률은 자외선 단독 처리군의 수준으로 증가되었다. 이는, 이 저해제가 MMS와 자외선으로 유발된 절제회복의 incision step에는 독립적으로 작용하며, 그 이후의 단계에서, MMS에 의해 부분적으로 불활성화 되었던 pyrimidine dimers의 절제를 완전하게 해주는 것으로 해석된다.

• 폴리머
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• 제35권5호
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• pp.431-437
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• 2011

아크릴계 이중인산과 과황산암모늄을 개시제로 사용하여 고분자 코팅용액의 기반인 poly(acryloyl diphosphoric acid)(poly(ADP))를 70 $^{\circ}C$의 수용액에서 라디칼 중합반응으로 제조하였다. 제조된 고분자 코팅용액인 poly(ADP)는 salmonella typhimurium, pseudomonas aeruginosa, escherichia coli, 그리고 staphylococcus aureus에 대한 항균성을 보였으며, 또한, Asperigillus niger와 조류인플루엔자 바이러스인 H1N1 바이러스에 대해서도 좋은 항진균성을 보여주었다. 더욱이 면섬유에 제조된 코팅용액 poly(ADP)를 코팅시켰을 경우에 우수한 난연성을 보여주었다.

• Journal of Magnetics
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• 제19권4호
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• pp.357-364
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• 2014

This study examined effect of a transcranial magnetic stimulation device with a commercial-frequency approach on the neuronal cell death caused ischemia. For a simple transcranial magnetic stimulation device, the experiment was conducted on an ischemia induced rat by transcranial magnetic stimulation of a commercial-frequency approach, controlling the firing angle using a Triac power device. The transcranial magnetic stimulation device was controlled at a voltage of 220 V 60 Hz and the trigger of the Triac gate was varied from $45^{\circ}$ up to $135^{\circ}$. Cerebral ischemia was caused by ligating the common carotid artery of male SD rats and reperfusion was performed again to blood after 5 minutes. Protein Expression was examined by Western blotting and the immune response cells reacting to the antibodies of Poly ADP ribose polymerase in the cerebral nerve cells. As a result, for the immune response cells of Poly ADP ribose polymerase related to necrosis, the transcranial magnetic stimulation device suppressed necrosis and had a protective effect on nerve cells. The effect was greatest within 12 hours after ischemia. Therefore, it is believed that in the case of brain damage caused by ischemia, the function of brain cells can be restored and the impairment can be improved by the application of transcranial magnetic stimulation.

• Molecules and Cells
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• 제21권2호
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• pp.218-223
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• 2006

The effect of poly(ADP-ribosyl)ation on the stability of p53 in SK-HEP1 cells treated with UV light was examined. Intracellular levels of p53 increased in cells treated with a low dose of UV light ($20J/m^2$), whereas they increased but then declined after a higher dose of UV ($100J/m^2$). Intracellular levels of p53 in the UV treated SK-HEP1 cells were dependent on the UV dose. Use of proteasome inhibitors revealed that p53 is degraded by proteasomal proteolysis after high doses of UV light. We present evidence that, at low doses, poly(ADP-ribose)polymerase (PARP) poly(ADP-ribosyl) ates p53 and protects it from proteasomal degradation before caspase-3 is activated, whereas at high doses the cells undergo UV induced apoptosis and PARP is cleaved by caspase-3 before it can protect p53 from degradation. Destabilization of p53 by cleavage of PARP may be important in cell fate decision favoring apoptosis.

• Molecules and Cells
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• 제37권1호
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• pp.9-16
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• 2014

ADP-ribosylation is a type of posttranslational modification catalyzed by members of the poly(ADP-ribose) (PAR) polymerase superfamily. ADP-ribosylation is initiated by PARPs, recognized by PAR binding proteins, and removed by PARG and other ADP-ribose hydrolases. These three groups of proteins work together to regulate the cellular and molecular response of PAR signaling, which is critical for a wide range of cellular and physiological functions.

• Bulletin of the Korean Chemical Society
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• 제33권4호
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• pp.1147-1153
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• 2012

Benzofuran-7-carboxamide was identified as a novel scaffold of poly(ADP-ribose) polymerase-1 (PARP-1) inhibitor. A series of compounds with various 2-substituents including (tertiary amino)methyl moieties substituted with aryl ring and aryl groups containing tertiary amines, were synthesized and biologically evaluated to elucidate the structure-activity relationships and optimize the potency. 2-[4-(Pyrrolidin-1-ylmethyl)phenyl]-benzofuran-7-carboxamide (42) was the most potent as an IC50 value of 40 nM among those.