• 농업과학연구
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• 제22권2호
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• pp.127-133
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• 1995

일련의 phenyl N-methylcarbamate 유도체들을 합성하고 ACh.E에 대한 carbamylation 과정으로 일어나는 살충활성 반응의 중심인 carbonvl group의 분자궤도(MO) 함수량과 살충활성($pI_{50}$) 값에 대한 치환기의 영향을 검토한 바, phenyl group에 결합된 치환기(X) 변화에 따른 carbonyl group의 알짜전하와 적외선 진동수($cm^{-1}$) 및 bond population에 미치는 치환기 효과(${\rho}=0$)는 관측되지 않았다. 전체 에너지(${\Delta}Et$)와 ipso $C_1-O_7$결합을 회전축으로 한 각도(${\theta}$)와의 관계로 부터 가장 안정한 형태는 평면인 phenyl group에 대하여 carbamyl group이 수직(${\theta}=90^{\circ}$ )인 (Z)-N-methyl-(Z}-O-phenylcarbamate 형태이었다. Carbamylation 과정의 1단계 평형($k_1/k_{-1}$) 반응은 thermodynamically control 반응이기 때문에 carronyl oxygene에 대한 acidic group(ACh.E)의 양성자화 반응과 Serine hydroxyl group의 친핵성 첨가반응 그리고 m-methyl group의 hyperconjugation effect에 의하여 anionic site(ACh.E)와의 상호작용으로 설명되는 일련의 새로운 carbamylation 메카니즘을 가정하여 제안하였다.

• KSBB Journal
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• 제11권3호
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• pp.295-302
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• 1996

Rubisco의 parameter들을 측정하는 것은 광합성 연구에 대단히 중요한데, 본 연구에서는 방사성통위 원소를 사용하는 방법 이 아닌 photometric assay 방 법을 사용하여 parameter를 측정하였다. Rubisco 의 activation과 carbamylation에 미치는 고 CO2 놓도의 효과를 연구하기 위하여, 정상 $CO_2$ 농도 (350 ppm)와 고 $CO_2$ 농도(650 ppm)에서 기른 kidney bean (Phaseolus vulgaris L.) 잎을 각각 재료로 하여, dual beam(334 nm9-} 405 nm) spectro­p photometer를 사용하여 rubisco의 initial activity 와 total activity, carbamylation rate 및 量을 측정 비교하였으며, SDS-PAGE에 의해 rubisco의 전기 영통 profile을 분석하였다. 정상 $CO_2$ 농도에서의 initial activity와 total activity는 $41.2{\mu}M/m^2/s 와 52.2{\mu}M/m^2/s$이며, 고 $CO_2$ 농도에서는 $27.4{\mu}M/m^2/s 와 46.1{\mu}M/m^2/s$로서, 350 ppm에서 650 ppm으로 $CO_2$ 농도를 증가시키면 rubisco의 initial activity와 total activity가 갑소되었다. 또한 carbamylation율도 정상 $CO_2$ 농도에서는 79%이며, 고 $CO_2$ 농도에서는 58.9%로서, $CO_2$ 농도의 증가에 따라 감소되 었다. Rubisco 의 量은 정상 $CO_2$농도에서는 $1.94 {\mu}M/m^2$$ 임에 반해 고 $CO_2$ 농도에서는 $1.58{\mu}M/m^2$ 로서, $CO_2$2 농도는 증가되었는데 그 量은 감소되었다. 이와 같이 고 $CO_2$농도에셔 rubisco의 activity가 감소되는 것은 rubisco의 carbamylation에 기인되는 것으로 생각된다. SOS-PAGE 분석에서 50kO 분자량의 large s subunit와 14.5 kO의 분자량을 가지는 small subu n mt를 동정하였는데, 고 $CO_2$ 농도와 정상 $CO_2$ 농도 의 50kD와 14.5 kO band의 intensity를 비교하면 두 구 사이에 큰 차이가 발견되지 않았다. 고 CO2 농도에서 정상 $CO_2$ 농도로 switch한 rubisco의 parameter는 정상 $CO_2$ 농도에서의 pa­r rameter와 거의 비슷한 값을 나타내였는데, 이는 350 ppm의 정상 $CO_2$ 농도에 의해 rubisco의 acti­v vity가 회복되었음을 의미한다.

• 대한의생명과학회지
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• 제20권2호
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• pp.96-102
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• 2014

Staphylococcal enterotoxin B (SEB) is bacterial toxin that induces the activation of immune cells. Because the inhibition of pro-inflammatory effect of SEB can resolve the inflammation, I determined the influence of functional or structural change of SEB on immune cells. The post translational modification of protein occurs through carbamylation. Carbamylation can change the structure of proteins and can modify the biological activity of protein. In the present study, I investigated the effect of carbamylated SEB (CSEB) on the inflammatory response mediated by LPS in HL-60 cells. To determine the anti-inflammatory effect of CSEB, I produced carbamylated SEB using potassium cyanate (KCN) and then examined whether CSEB involved in cytokine releases and apoptosis of LPS-stimulated HL-60 cells. Although CSEB had not any effect on the LPS-stimulated HL-60 cells, the protein levels of IL-8, TNF-${\alpha}$ and IL-$1{\beta}$ were significantly decreased by CSEB without cytotoxicity. CSEB also blocked Akt and NF-${\kappa}B$ activation. These results indicate that the suppressive effect of CSEB in LPS-stimulated cytokine releases is occurred by inhibition of Akt and NF-${\kappa}B$ activity. Through further studies, CSEB may be used as anti-inflammatory molecule that makes the immune system more efficient.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1994년도 한국생물과학협회 학술발표대회
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• pp.190.1-190
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• 1994

• Applied Biological Chemistry
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• 제32권2호
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• pp.170-177
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• 1989

2개의 m-methyl group이 치환된 m-xylyl N-methylcarbamates(MXNMC)가 비치환된 phenyl N-methylcarbamates(PNMC)보다 높은 살충작용을 나타내는 이유를 알아보기 위하여 이들 두 유도체들이 확장 $H\ddot{u}ckel$(EHT) 이론에 따른 분자궤도(MO) 이론과 회귀 및 자유에너지 관계(LFER) 분석에 의하여 연구되었다. 가장 안정한 입체구조는 phenyl group이 N-methylcarbamyl group의 면에 대하여 수직(${\theta}=90^{\circ}$)인 형태(Z,Z)이었으며, 회귀분석은 $pI_{50}$ 상수들과 meta-위치 및 m-methyl group의 수소원자 하전과 LUMO에너지의 계산된 MO량을 변수로 취했을 경우가 가장 좋은 상관관계를 나타내었다. Carbamylation 반응에 관한 LFER분석 결과, PNMC에서는 공명(R)-효과(40%)보다 장 (F) 효과(60%)가 약간 컸으나(F>R), MXNMC의 경우에는 R-효과(98.6%)가 F-효과(1.4%)보다 월등히 큰 값($R{\gg}F$)을 보였다. 이와 같은 사실로부터 MXNMC의 살충작용이 증진되는 것은 m-dimethyl group에 의한 hyperconjugation의 결과라고 믿어진다.

• 한국방사선학회논문지
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• 제13권1호
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• pp.125-132
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• 2019

Potassium cyanate는 무기화합물로 단백질의 번역 후 과정에서 카바밀화(carbamylation)을 유도할 수 있고 이러한 카바밀화 반응은 다양한 질병 및 조건에서 세포의 사멸과 관련이 있다. 이전 연구결과에서 KCN은 사람 대장암 세포주인 HCT 116세포의 방사선 감수성을 향상시키는 것을 확인하였지만 그 기전을 명확히 규명하기에는 많이 부족한 실정이다. 본 연구에서는 방사선에 다소 저항성을 가지는 대장암 세포에서 KCN이 방사선 감수성을 향상시키고 세포사멸 시키는 기전을 확인하기 위해 2 mM의 KCN 처리 후 저 선량의 광자선을 조사하여 세포주기, 세포 생존율, 세포 사멸 관련 단백질(caspase-1, PARP) 발현량, $TNF-{\alpha}$ 분비 및 $TNF-{\alpha}$ 관련 전사인자($NF-{\kappa}B$)의 연관성을 확인하였다. 그 결과 KCN 처리 후 광자선을 조사한 세포에서 caspase-3 및 PARP의 활성이 증가하고 이는 세포주기의 정지와 세포사멸을 유도하였다. 또한 이 과정에서 DNA 전사인자인 $NF-{\kappa}B$에 의해 세포 외로 $TNF-{\alpha}$를 지속적으로 분비하여 세포사멸에 관여함을 확인하였다. 이러한 결과들을 토대로 KCN이 radiosensitizer로서 작용할 수 있는 가능성이 있다고 사료된다.

• 대한의생명과학회지
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• 제19권2호
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• pp.98-104
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• 2013

Radiation is one of the major therapy for the removal of cancer cells. The results of the radiation therapy depend on the radio-resistance of cancer cells. For the effective treatment in these radio-resistant cancers, the use of chemicals that act on cancer cells is known to enhance the cytotoxic effects of radiation therapy. In this study, I investigated the effect of potassium cyanate (KCN) on the irradiated-colorectal cancer cell line, HCT 116 cells. KCN induces the carbamylation of proteins and can change the biological activity of various human cells. To understand the effect of KCN on the radiosensitivity of HCT 116 cells, I examined alteration of the cell cycle, generation of reactive oxygen species (ROS), cell viability, apoptosis and intracellular signaling proteins in the irradiated cells with/without KCN treatment. Combination treatment caused significant increase in sub $G_0/G_1$ and ROS generation in HCT 116 cells. KCN inhibited the proliferation and cell viability in irradiated HCT 116 cells. KCN-induced apoptosis of irradiated cells was processed via the activation of caspase 3 and caspase 9. Apoptosis-associated signal proteins, including Bax and Bcl-2 were regulated by irradiation with KCN treatment. Taken together, these results may indicate that KCN enhances the radiosensitivity of radio-resistant cell and then has a synergistic effect on radiation therapy in colorectal cancer.

• 대한임상독성학회지
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• 제5권1호
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• pp.74-78
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• 2007

The carbamates are a group of insecticides derived from carbamic acid, with a broad spectrum of uses as agricultural and household garden insecticides. Carbamate insecticides are reversible cholinesterase inhibitors. Their inhibitory action is mediated by reversible carbamylation of acetylcholine, as with the organophosphate insecticides. Carbamates are absorbed by the body through multiple routes, including inhalation, ingestion, and dermal absorption. Although poisoning can result from occupational exposure or accidental ingestion, in most cases there is suicidal intent. This is particularly true in developing countries, where the highest incidence of morbidity and mortality from this cause occurs. Cardiac complications often accompany poisoning by carbamate compounds, which may be serious and often fatal. The extent, frequency, and pathogenesis of cardiac toxicity from carbamate compounds has not been clearly defined. Possible mechanismsinclude sympathetic and parasymphatetic overactivity, hypoxemia, acidosis, electrolyte derangements, and a direct toxic effect of the compounds on the myocardium. Patients with carbamate poisoning should immediately be transferred to an intensive or coronary care unit where appropriate monitoring and resuscitative facilities are available. We here report a case of acute coronary syndrome resulting from acute carbamate ingestionthat resulted in a healthy discharge.

• 대한의생명과학회지
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• 제17권3호
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• pp.177-184
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• 2011

Potassium cyanate (KOCN) is an inorganic compound and induces the carbamylation of proteins with cytotoxic effects on human cells. Although there is a potential cytotoxic molecule, the role of KOCN on the apoptosis of cancer cell is not well understood. The present study investigated the effects of KOCN on the human colorectal cancer cell line, HCT 116 cells. To understand the anti-cancer effect of KOCN on HCT 116 cells, we examined alteration of apoptosis, the intracellular $Ca^{2+}$ concentration, the intracellular signaling pathway and generation of reactive oxygen species (ROS) in these cells treated with KOCN. The apoptosis of HCT 116 cells was induced by KOCN in a dose-dependent manner at 24 hours and 48 hours, respectively. The apoptosis was processed via the cleavage of poly ADP-ribose polymerase (PARP) and activation of caspase 3 in HCT 116 cells. KOCN induced the elevation of intracellular $Ca^{2+}$ concentration and changed the expressions of Bcl-2 family proteins. The pro-apoptotic Bax was continuously up-regulated, and the anti-apoptotic Bcl-2 was down-regulated by KOCN. KOCN also induced the hyperpolarization of mitochondria and the generation of ROS in HCT 116 cells. Taken together, these results indicate that KOCN induces the apoptosis of HCT 116 cells by disruption of $Ca^{2+}$ homeostasis and via mitochondrial pathway. This study provides the compound that may be used as a potent agent for the treatment of colorectal cancer.

• 대한의생명과학회지
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• 제20권1호
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• pp.32-38
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• 2014

Potassium cyanate (KOCN) that is known as an inducer of the protein carbamylation is an inorganic compound and is the conjugate based of cyanic acid (HOCN). Based on these studies, we confirmed that KOCN induces the apoptosis of the human colorectal cancer cell line, HCT 116 cells, by various mitochondrial pathways. To investigate other mechanisms of KOCN-mediated apoptosis, in the present study, we examined KOCN-induced cytokines production in HCT 116 cells and identified the intracellular signaling pathway in these processes. We first demonstrated that KOCN considerably increased the cell apoptosis via intracellular $Ca^{2+}$ signaling, mitochondrial dysfunction and ROS production. And then we examined TNF-${\alpha}$ and IL-$1{\beta}$ levels mediated by KOCN in HCT 116 cells. Although IL-$1{\beta}$ was not involved in KOCN-mediated HCT 116 cell apoptosis, the release of TNF-${\alpha}$ was mediated by KOCN in HCT 116 cells via NF-${\kappa}B$ activation. Apoptosis was also enhanced by incubation with supernatants from HCT 116 cells after KOCN treatment and this effect was partially reduced by BAY 11-7085 pre-treated supernatant. Taken together, our results indicate that KOCN-induced apoptosis in HCT 116 cells is dependent on the releases of TNF-${\alpha}$ and the increased factors and that the mechanism involves the activation of NF-${\kappa}B$.