• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1275-1280
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• 2015

Previously, we performed de novo RNA sequencing of Scolopendra subspinipes mutilans using high-throughput sequencing technology and identified several antimicrobial peptide candidates. Among them, a cationic antimicrobial peptide, scolopendrasin VII, was selected based on its physicochemical properties, such as length, charge, and isoelectric point. Here, we assessed the anticancer activities of scolopendrasin VII against U937 and Jurkat leukemia cell lines. The results showed that scolopendrasin VII decreased the viability of the leukemia cells in MTS assays. Furthermore, flow cytometric analysis and acridine orange/ethidium bromide staining revealed that scolopendrasin VII induced necrosis in the leukemia cells. Scolopendrasin VII-induced necrosis was mediated by specific interaction with phosphatidylserine, which is enriched in the membrane of cancer cells. Taken together, these data indicated that scolopendrasin VII induced necrotic cell death in leukemia cells, probably through interaction with phosphatidylserine. The results provide a useful anticancer peptide candidate and an efficient strategy for new anticancer peptide development.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.2046-2056
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• 2018

Phospholipase D has great commercial value due to its transphosphatidylation products that can be used in the food and medicine industries. In order to construct a strain for use in the production of PLD, we employed a series of combinatorial strategies to increase PLD expression in Bacillus subtilis WB600. These strategies included screening of signal peptides, selection of different plasmids, and optimization of the sequences of the ribosome-binding site (RBS) and the spacer region. We found that using the signal peptide amyE results in the highest extracellular PLD activity (11.3 U/ml) and in a PLD expression level 5.27-fold higher than when the endogenous signal peptide is used. Furthermore, the strain harboring the recombinant expression plasmid pMA0911-PLD-amyE-his produced PLD with activity enhanced by 69.03% (19.1 U/ml). We then used the online tool \RBS Calculator v2.0 to optimize the sequences of the RBS and the spacer. Using the optimized sequences resulted in an increase in the enzyme activity by about 26.7% (24.2 U/ml). In addition, we found through a transfer experiment that the retention rate of the recombinant plasmid after 5 generations was still 100%. The final product, phosphatidylserine (PS), was successfully detected, with transphosphatidylation selectivity at 74.6%. This is similar to the values for the original producer.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.578-589
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• 2003

The fuzzy rat that expresses hypersecretion of sebum and hyperplastic sebaceous glands is a genetic mutant for the study of many pharmacological aspects especially human acne. Through this model, we examined the effects of several phospholipids on the secretion of sebum after topical application. The phospholipid derivatives were phosphatidylcholine (PC), hydrogenated phosphatidylcholine (HPC), phosphati dylserine (PS) and hydrogenated phosphatidylserine(HPS). All agents were dissolved into the vehicle (1, 3-Butanediol, ethanol and water) at 0.5％ weight volume and applied on the dorsal area of the fuzzy rat. To observe histological changes, the skin biopsies were stained with Oil Red O and the size and morphology of sebaceous gland was observed under microscope. Topical treatment with PC and/or HPC showed a marked decrease in sebum excretion. Especially hydrogenated PC (HPC) appeared to have more predominant sebosuppressive function than any other treatment. The other agents such as PS and HPS showed a marginal effect on sebum secretion. With the sebosuppressive activity, HPC and PC seem to have a good potential application on acne treatment. In order to obtain more insights into possible mechanisms behind the above observations, effects of each phospholipid on the expression of peroxisome proliferator-activated receptor (PPAR) genes were investigated. Recently, it has been demonstrated that expression and activation of PPAR subtypes appear to modulate the accumulation of cytoplasmic fat droplets that characterizes the sebocyte differentiation(1). It was also previously suggested that PPAR${\gamma}$ antagonist would seem possible to interfere sebum production without side effects (2). In this study we examined the diverse effects of the tested phospholipids on the expression of several PPAR genes based on reverse transcription-polymerase chain reaction (RT-PCR) from the topically treated skin of fuzzy rats. The results and possible implications are discussed.

• KSBB Journal
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• 제7권4호
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• pp.302-307
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• 1992

protein phosphatase 2A는 bovine brain homogenate의 세포질 fraction에서 얻어졌다. 기질로서 인산화된 histione H1을 이용하여 측정한 phosphatase 의 활성은 dipalmitoyIphophatidylcholine(DPPC) 혹은 phosphatidylserine/DPPC의 혼합물로 구성된 liposome의 존재하에서 저해되었다. Protein phosphatase 2A의 lipid membrane에의 결합은 다중층 지질막의 혼합물 계에서 liposome 의 양이 증가함에 따라서 상등액 중의 phosphatase의 활성이 감소하는 것으로 확인할 수 있었다. 또한 [$^{125}I$]protein phosphatase 2A가 liposome과 동시에 용출되는 것으로도 확인되었다. 그러나 liposome에 대한 protein phosphatase의 친화력은 높지 않았다. 한편, okadaic acid와 liposome은 협동으로 phosphatase의 활성을 감소시켰다. 이것은 okadaic acid가 lipid membrane이나 membrane에 결함한 phosphatase에는 결합하지 않는다는 것을 의미한다. 그러므로 lipid membrane에 의한 protein phosphatase 2A의 활성 저해 효과는 phosphatase 2A와 lipid membrane과의 결합에 의한 것이라고 설명될 수있다.

• 약학회지
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• 제48권2호
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• pp.153-158
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• 2004

Paecilomyces tenuipes is one of the famous Chinese medicinal entomopathogenic fungi that parasite in the lavae of silkworm. A cytotoxic compound, 4$\beta$-acetoxyscirpendiol (ASD) was isolated from a methanolic extracts of Paecilomyces tenuipes. The ASD compound belongs to scirpenol subfamily of trichothecene mycotoxin. In a continuation of the elucidation of the mechanism of ASD, we report here the evidences of induction of apoptosis by ASD in human Jurkat T cell line. In MTT reduction assay for monitoring cell viability, ASD showed strong toxicity. The 50 percent inhibitory concentrations of ASD against human T lymphoid Jurkat cell was 59.5 ng/$m\ell$. Phosphatidylserine externalization was increased by ASD at 3 and 6 hrs when compared with that of 6 hrs in the cell line showing in a time-dependent manner. When whole lysates of cells treated with ASD were subjected to western blot assay, 113 kDa poly(ADP-ribose) polymerase (PARP) was significantly cleaved to 89 kDa fragment. Time-dependent DNA fragmentation was also observed when Jurkat T cells were treated with ASD at 100 ng/$m\ell$ for 6 hrs and 18 hrs at the ratios of 8.5% and 15.0%, respectively. From these data, Jurkat T lymphocytes treated with ASD from Paecilomyces tenuipes underwent typical cascades of apoptotic cell death.

• Toxicological Research
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• 제22권4호
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• pp.397-402
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• 2006

Lipid mediator such as lysophosphatidic acid (LPA) plays an important role in inflammation and wound heating, has been recently reported to induce influx of extracellular calcium into erythrocytes. This elevation in intracellular calcium level may cause destruction of membrane asymmetry and procoagulant microvesicle formation. Thus, we investigated if the lipid mediator could induce microvesicle formation as a result of extracellular calcium influx in human erythrocytes. Treatment with lipid mediator to erythrocytes resulted in microvesicle generation In a concentration-, time-dependent manner. Microvesicles formed expressed procoagulant phosphatidylserine (PS) on their surface membrane significantly as well. LPA did not affect the band 3 phosphorylation which is involved in morphological change in erythrocytes. Pretreatment with suramin did not inhibit LPA-induced microvesicle generation, suggesting microvesicle generation was not receptor-dependent pathway. Depletion of intracellular ATP levels in erythrocytes was suggested to be one of the mechanism for these events.

• 한국균학회지
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• 제34권2호
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• pp.119-121
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• 2006

키토산및 올리고키토산의 C. albicans과 T. rubrum에 대한 항 진균작용에 대하여 조사하였다. 키토산은 C. albicans과 T. rubrum에 대하여 항 진균 효과를 나타내었으나 올리고키토산은 두 균주에 대한 성장억제를 나타내지 않았다. 키토산의 항 진균 양상은 살 진균 적 활성을 나타내었다. 키토산의 apoptosis유도에 대한 조사를 위하여 phosphatidylserine의 세포막 외부로의 노출을 FITC-Annexin 반응을 이용하여 관찰하였다. 실험결과 키토산은 C. albicans의 세포에 대하여 apoptosis를 유도하는 것을 보여주었다.

• 동의생리병리학회지
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• 제28권3호
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• pp.337-345
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• 2014

The present study examined the effects of soybean-derived phosphatidylserine (SB-PS) on the learning and memory function and the neural activity in rats with trimethyltin (TMT)-induced memory deficits. The cognitive improving efficacy of SB-PS on the amnesic rats, which was induced by TMT, was investigated by assessing the Morris water maze test and by performing cholineacetyl transferase (ChAT), acetylcholinesterase (AChE) and cAMP responsive element binding protein (CREB) immunohistochemistry. A positron emission tomography (PET) scanning the rat brain was by performed administer 18F-Fluorodeoxy-glucose (18F-FDG). The rats with TMT injection showed impaired learning and memory of the tasks and treatment with SB-PS produced a significant improvement of the escape latency to find the platform in the Morris water maze at the 2nd day compared to that of the MCT group. In the retention test, the SB-PS group showed increased time spent around the platform compared to that of the MCT group. Consistent with the behavioral data, SB-PS 50 group significantly alleviated the loss of acetyl cholinergic neurons in the hippocampus compared to that of the MCT group. Treatment with SB-PS significantly increased the CREB positive neurons in the hippocampus as compared to that of the MCT group. In addition, SB-PS groups increased the glucose uptake in the hippocampus and SB-PS 50 group increased the glucose uptake in the frontal lobe, as compared to that of the MCT group. These results suggest that SB-PS may be useful for improving the cognitive function via regulation of cholinergic marker enzyme activity and neural activity.

• 대한약리학회지
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• 제30권1호
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• pp.117-124
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• 1994

Phospholamban은 심근 근장그물 $Ca^{2+}-ATPase$ 조절단백이다. 조절작용기전은 탈인산화된 phospholamban에 의해 $Ca^{2+}-ATPase$가 억제됨으로 나타나며, 이 phospholamban이 인산화됨으로 $Ca^{2+}-ATPase$에 대한 억제가 반전됨을 보인다. 최근에 phospholamban의 cytoplasmic domain만으로 $Ca^{2+}-ATPase$를 억제하기에는 불충분하다는 보고가 있어 본 실험을 계획하였다. $Ca^{2+}-ATPase$의 활성을 억제하는 phospholamban domain을 밝히기 위하여 합성한 phospholamban 펩타이드(아미노산 1-25)의 $Ca^{2+}$ uptake에 대한 효과를 살펴보았다. 골격근 근장그물에서 $Ca^{2+}-ATPase$를 분리한 후 phosphatidylcholine이나 phosphatidylcholine과 phosphatidylserine을 포함한 liposome에 재조합시켰다. Phospholamban 펩타이드는 phosphatidylcholine을 이용하여 재조합된 vesicles의 초기 $Ca^{2+}$ uptake rate를 억제하고, cAMP 의존성 protein kinase의 catalytic subunit로 인산화시킨 phospholamban 펩타이드는 이 억제를 반전시킴을 보여 주었다. Phosphatidylcholine과 phosphatidylserine을 포함한 제조합 vesicles에서도 같은 양상을 보였다. 이상의 결과로 미루어 볼 때 인산화 sites를 포함하고 있는 phospholamban의 cytoplasmic domain은 그 자체만으로도 근장그물 칼슘펌프를 억제하기에 충분하다고 결론지을 수 있다.

• 대한화장품학회지
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• 제32권1호
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• pp.17-22
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• 2006

포스파티딜세린(Phosphatidylserine; PS)은 생체막에서 구조적인 역할을 담당하는 인지질로서, 생체 내 다양한 세포작용에 필수적인 신호전달 효소의 보조인자로서 작용하는 것으로 알려져 있다. 하지만, PS의 생리활성에 대한 연구는 거의 이루어지지 않았고, 특히 피부에서의 생리활성에 대한 연구는 전무한 실정이다. 본 연구에서는 무모생쥐의 피부에 tape-stripping으로 경표피수분손실(TEWL)의 증가를 유도한 후, PS를 도포함으로써 그 손실을 현저히 감소시켰다. 또한, PS 도포군의 피부에서 세라마이드 함량이 증가된 사실을 확인한 바 있다. PS 도포군에서 non-hydroxyl 세라마이드와 glucosyl 세라마이드의 함량이 비처리군과 비교하여 각각 1.4배와 1.6배로 증가하였다. PS는 또한 피부각질세포의 분화를 촉진하였다. 피부각질세포에 PS를 처리함으로써 세포 형태가 분화상을 띄고 있음을 현미경 상에서 확인하였고, 표피분화의 특이적 표지 단백질인 Involucrin (INV)과 Transglutaminase 1 (TG'ase 1)의 발현이 각각 3.5배와 3배로 현저히 증가하였음을 웨스턴 블랏을 통하여 확인하였다. 또한 무모생쥐 피부에 PS를 도포한 결과 INV와 loricrin 단백질 발현이 증가하였다. 본 연구는 PS가 피부에서 생리활성을 나타낸다는 최초의 증거를 제시하며, 구체적으로는 각질세포 분화를 촉진함으로써 피부 세라마이드 함량을 증가시키고 경표피 수분손실을 감소시켜 궁극적으로 피부장벽을 강화하는 작용을 한다는 것을 보여준다.