• 한국미생물·생명공학회지
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• 제15권6호
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• pp.425-429
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• 1987

B. thuringiensis subsp. israelensis 균주가 생성하는 $\delta$-endotoxin의 alkali 용해에서 생성되는 hemolysin fragment 와 extra-cellular hemolysin 의 상관관계를 조사하기 위한 기초자료로서 extra-cellular hemolysin을 분리 정제하여 그 성질을 조사하였다. 균체 배양액에 유안을 염석 및 투석시킨 후, Sephadex G-200 gel filtration 과 DEAE-cellulose column chromatography를 행하여 균체외 homely-sin을 정제하였다. 정제된 extra-cellular hemolysin 은 SDS-polyacrylamide gel 전기영동에서 47,000 dalton의 분자량을 가진 단일 단백질 band를 얻었다. 또한 정제된 hemolysin은 thiol agents에 의해 그 활성이 증가되었으나, cholesterol 및 protease 처리 또는 금속이온 즉, FeSO$_4$나 CuSO$_4$에 의해 그 활성 이 감소되었다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.127-127
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• 1998

Specific inhibitors of a calcium activated neutral protease calpain could be used for the treatment of neurodegenerative diseases, cataract and muscular dystrophy diseases because of their therapeutic effects. In the course of screening for potential calpain inhibitors from microorganisms, a new analogue of chymostatins named calmostinol was isolated from the culture filtrate of an actinomycete. The MW was determined to be 596 [(M + H)$\^$+/] by FAB-MS in glycerol matrix. The structure was elucidated to be N-[((S)-1-carboxy-2-phenylethyl)-carbamoyl]-${\alpha}$-[2- iminohexahydro-4(S)-pyrimidyl]-L-glycyl- L-valyl-phenylalaninol, by the spectroscopic methods such as NMR and MS fragmentation studies. Calmostinol exhibited strong activity against calpain while not against a Ca$\^$2+/ -independent cysteine protease papain.

• 대한약침학회지
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• 제3권1호
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• pp.1-19
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• 2000

To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability and apoptosis were analyzed using MTT assay, tryphan blue assay, $[^3H]$thymidine release assay, flow cytometric analysis, and activity of caspase-3 protease activity assay. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, $[^3H]$thymidine release assay, and flow cytomet1 ric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and Bcl-$X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment.

• Journal of Korean Neurosurgical Society
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• 제30권sup1호
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• pp.5-12
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• 2001

This study was designed to investigate the mechanism of cell death after cisplatin treatment in human glioblastoma A172 cells. Cis-diamminedichloroplatinum(Cisplatin) demonstrated cytostatic or cytotoxic effects on A172 cells in a dosedependent manner. Cisplatin-mediated cytotoxity in A172 cells was revealed as an apoptosis characterized by high molecular weight DNA fragmentation by agarose electrophoresis as well as nuclear fragmentation by Hoechst staining. Cisplatin also resulted in the activation of caspase 3-like protease as well as poly(ADP-ribose) polymerase(PARP) cleavage. Interestingly, the anti-apoptotic Bcl2 protein was degraded and furthermore, expression of p53 protein was increased by cisplatin in a time-dependent manner. Taken together, these results suggest that anticancer drug, cisplatin induces the apoptotic death of human glioblastoma A172 cells via the activations of caspase 3-like protease, degradation of anti-apoptotic Bcl2 protein and increase in the expression of p53.

• BMB Reports
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• 제56권8호
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• pp.451-456
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• 2023

Deubiquitinases (DUBs) are an essential component of the ubiquitin-proteasome system (UPS). They trim ubiquitin from substrate proteins, thereby preventing them from degradation, and modulate different cellular processes. Ubiquitin-specific protease 14 (USP14) is a DUB that has mainly been studied for its role in tumorigenesis in several cancers. In the present study, we found that the protein levels of USP14 were remarkably higher in gastric cancer tissues than in the adjacent normal tissues. We also demonstrated that the inhibition of USP14 activity using IU1 (an USP14 inhibitor) or the inhibition of USP14 expression using USP14-specific siRNA markedly reduced the viability of gastric cancer cells and suppressed their migratory and invasive abilities. The reduction in gastric cancer cell proliferation due to the inhibition of USP14 activity was a result of the increase in the degree of apoptosis, as evidenced by the increased expression levels of cleaved caspase-3 and cleaved PARP. Furthermore, an experiment using the USP14 inhibitor IU1 revealed that the inhibition of USP14 activity suppressed 5-fluorouracil (5-FU) resistance in GC cells. Collectively, these findings indicate that USP14 plays critical roles in gastric cancer progression and suggest its potential to serve as a novel therapeutic target for gastric cancer treatment.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2020년도 추계국제학술대회
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• pp.14-14
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• 2020

In December 2019, the COVID-19 epidemic was discovered in Wuhan, China, and since has disseminated around the world impacting human health for millions. Herein, in-silico drug discovery approaches were utilized to identify potential candidates as Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) main protease (M^pro) inhibitors. We investigated several databases including natural and natural-like products (>100,000 molecules), DrugBank database (10,036 drugs), major metabolites isolated from daily used spices (32 molecules), and current clinical drug candidates for the treatment of COVID-19 (18 drugs). All tested compounds were prepared and screened using molecular docking techniques. Based on the calculated docking scores, the top ones from each project under investigation were selected and subjected to molecular dynamics (MD) simulations followed by molecular mechanics-generalized Born surface area (MM-GBSA) binding energy calculations. Combined long MD simulations and MM-GBSA calculations revealed the potent compounds with prospective binding affinities against M^pro. Structural and energetic analyses over the simulated time demonstrated the high stabilities of the selected compounds. Our results showed that 4-bis([1,3]dioxolo)pyran-5-carboxamide derivatives (natural and natural-like products database), DB02388 and Cobicistat (DB09065) (DrugBank database), salvianolic acid A (spices secondary metabolites) and TMC-310911 (clinical-trial drugs database) exhibited high binding affinities with SARS-CoV-2 M^pro. In conclusion, these compounds are up-and-coming anti-COVID-19 drug candidates that warrant further detailed in vitro and in vivo experimental estimations.

• 동의생리병리학회지
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• 제18권3호
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• pp.759-766
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• 2004

In the present study, we investigated the effects of aqueous extract of the healthful decoction utilizing Phellinus linteus (HDPL) on the cell growth of human lung carcinoma tumor cell line A549. Exposure of A549 cells to HDPL resulted in growth inhibition and induction of apoptosis in a dose-dependent manner as measured by hemocytometer counts, fluorescence microscopy and flow cytometric analysis. This increase in apoptosis was associated with inhibition and/or degradation of apoptotic target proteins such as poly(ADP-ribose) polymerase (PARP), b-catenin and phospholipase C- 1 (PLC- 1) protein. HDPL treatment induced the down-regulation of anti-apoptotic Bcl-2 expression, an anti-apoptotic gene, however, the level of Bax. a pro-apoptotic gene, was increased by HDPL treatment. In addition, HDPL-induced apoptotis of A549 cells was connected with activation of caspase-3 and caspase-9 protease in a dose-dependent manner, however, the levels of inhibitor of apoptosis proteins family were remained unchanged. Taken together, these results indicated that the anti-proliferative effects of HDPL were associated with the induction of apoptotic cell death through regulation of several major growth regulatory gene products such as Bcl-2 family expression and caspase protease activity, and HDPL may have therapeutic potential in human lung cancer.

• 한국식품과학회지
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• 제28권4호
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• pp.678-683
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• 1996

Pronase 등 8개의 단백분해효소를 사용하여 말쥐치를 가수분해시 생성되는 펩타이드의 양을 측정한 결과, bromelain과 neutrase에 의한 가수분해물의 펩타이드 생성량은 4시간 가수분해시 6.13 mg/ml와 6.01 mg/ml로 높은 생성량을 보였으며, 유리아미노산의 생성량은 4시간 가수분해시 alcalase에 의한 가수분해물이 4.20 mg/ml로 높은 유리 아미노산의 생성량을 보였다. 가수분해도를 측정한 결과, 4시간 가수분해시 esp/sav와 alcalase의 가수분해물은 88.9%와 86.2%의 비교적 높은 가수분해도를 보였다. 또한 핵산관련 물질은 5'-GMP가 다른 핵산 관련물질에 비해 높은 생성량을 보였다. 가수분해물의 펩타이드의 평균길이는 $8.5{\sim}14.5$로 bromelain에 의한 가수분해물이 가장 짧은 펩타이드 길이를 가진 반면, neutrase에 의한 가수분해물은 14.5로 가장 긴 펩타이드로 구성되었다.

• Applied Biological Chemistry
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• 제47권2호
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• pp.197-201
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• 2004

식품 가공의 부산물 중 동물성 부산물을 재활용하여 조미소재로의 개발 가능성을 탐색하였다. 부산물 원료로는 돼지뼈(PB)와 닭뼈(CB), 그리고 참치 혈합육(TDF)을 사용하였다. PB와 CB를 열수로 18시간 추출한 후 단백질과 지방 분해효소를 처리하여 가수분해물을 제조하였다. 각각의 열수 추출물에서 지방의 제거를 위하여 1차로 lipase활성이 있는 pancreatic enzyme을 처리하고 2차로 Flavourzyme을 사용하는 경우 가수분해도(DH)가 PB는 약 70%, CB는 약 80%까지 증가하였다. 지방이 적은 TDF는 단백질 분해효소인 Alcalase와 Flavourzyme으로 분해한 결과 고형분 회수율은 약 22%, 단백질 회수율은 9%이었으며, 효소분해액 중 유리 아미노산 함량은 고형분 중 27%까지 향상되었다. 세 종류 분해액에 가염한 후 관능적인 특성을 조사한 결과 전체적으로 TDF>PB>CB의 순으로 우수하여 참치 혈합육은 조미소재의 원료로 적합한 동물성 부산물로 판단되었다.

• Biomolecules & Therapeutics
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• 제7권3호
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• pp.271-277
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• 1999

Safety evaluation of LB71350, a new HIV-1 protease inhibitor, was performed in mice, rats and dogs. For the general behavior of mice, LB71350 at an oral dose of 200 mg/kg did not show any significant effects on muscle tone and locomotor activity. In terms of central nervous system, at oral doses of 200 mg/kg and 1000 mg/kg, LB71350 inhibited acetic acid-induced pain response approximately 41% and 83% of control. respectively. At oral doses of 200 mg/kg and 500 mg/kg, it reduced the rectal body temperature in rats. Pentylenetetrazole-induced seizure in mice was slightly potentiated by oral administration of LB71350 at doses ranging from 200 mg/kg to 1000 mg/Ag. Single or five day treatment of LB71350 doubled the hexobarbital- induced sleeping time in mice at oral doses ranging from 50 mg/kg to 500 mg/kg. It did not cause any effects on gastric secretion and acidity in rat at oral doses of 200 mg/kg and 1000 mg/kg and also it did not change intestinal motility in mice up to 1000 mg/kg. Blood coagulation indices such as prothrombin time (PT), activated partial thromboplastin time (aPTT), and thrombin time (TT) in rats were not affected by the treatment of LB71350 up to 500 mg/kg. LB71350 caused no significant effects on the cardiac output, stroke volume, heart rate, and mean blood pressure when infused intravenously to the anesthetized rats and dogs. Taken together, LB71350 at high oral doses caused significant pharmacological effects on the central nervous system and the hexobarbital-induced sleeping time.