• Journal of Plant Biotechnology
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• 제49권4호
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• pp.316-324
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• 2022

Hair loss causes psychological stress due to its effect on appearance. Therefore, the global market for hair loss treatment products is rapidly growing. The present study demonstrated that ginseng berry-derived and sequence-modified peptides promoted the proliferation rate of dermal papilla (DP) cells and keratinocytes, in addition to having antioxidant properties. Moreover, the potential role of these ginseng berry peptides as TGF-β2 antagonists was confirmed through in silico computer docking. In addition to promoting the growth of ,the ginseng berry-derived peptides also promoted the proliferation of keratinocytes experimental Particularly, an unmodified ginseng berry-derived peptide (GB-1) and two peptides with sequence modifications (GB-2 and GB-3) decreased ROS generation and exhibited a protective effect on damaged HaCaT keratinocytes. Computer-aided peptide discovery was conducted to identify the potential interactions of important proteins with transforming growth factor-beta 2 (TGF-β2), a key protein that plays a crucial role in the human hair growth cycle. Our results demonstrated that MAGH, an amino acid sequence present in herbal supplements and plant-based natural compounds, can inhibit TGF-β2.

• Asian Pacific Journal of Cancer Prevention
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• 제16권5호
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• pp.1833-1837
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• 2015

An aristolactam-type alkaloid, isolated from Orophea enterocarpa, is enterocarpam-III (10-amino-2,3,4,6-tetramethoxyphenanthrene-1-carboxylic acid lactam). It is cytotoxic to various human and murine cancer cell lines; however, the molecular mechanisms remain unclear. The aims of this study were to investigate cytotoxic effects on and mechanism (s) of human cancer cell death in human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells compared to normal murine fibroblast NIH3T3 cells. Cell viability was determined by MTT assay to determine $IC_{10}$, $IC_{20}$ and $IC_{50}$ levels, reactive oxygen species (ROS) production with 2',7'-dichlorohydrofluorescein diacetate and the caspase-3, -8 and -9 activities using specific chromogenic (p-nitroaniline) tetrapeptide substrates, viz., DEVD-NA, IETD-NA and LEHD-NA and employing a microplate reader. Mitochondrial transmembrane potential (MTP) was measured by staining with 3, 3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and using flow cytometry. The compound was cytotoxic to HepG2 and MDA-MB-231 cells with the $IC_{50}$ levels of $26.0{\pm}4.45$ and $51.3{\pm}2.05{\mu}M$, respectively. For murine normal fibroblast NIH3T3 cells, the $IC_{50}$ concentration was $81.3{\pm}10.1{\mu}M$. ROS production was reduced in a dose-response manner in HepG2 cells. The caspase-9 and -3 activities increased in a concentration-dependent manner, whereas caspase-8 activity did not alter, indicating the intrinsic pathway activation. Enterocarpam-III decreased the mitochondrial transmembrane potential (MTP) dose-dependently in HepG2 cells, suggesting that the compound induced HepG2 cell apoptosis via the mitochondrial pathway. In conclusion, enterocarpam-III inhibited HepG2 and MDA-MB-231 cell proliferation and induced human HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway and induction of caspase-9 activity.

• KSBB Journal
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• 제25권4호
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• pp.395-400
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• 2010

본 연구에서는 파파인으로 가수분해된 피브로인 펩타이드에 글리세롤을 추가하고 동일한 효소인 파파인을 이용하여 글리세릴 에스터 결합을 형성시키는 반응을 수행하였다. 피브로인 펩타이드 10%와 글리세롤 50% 포함된 반응용액을 pH 3, $40^{\circ}C$의 반응 조건에서 글리세릴 에스터가 생성됨을 ESI 질량 분석을 통하여 확인하였다. 아울러 반응전 시료의 펩타이드의 조성과 생성된 글리세릴 펩타이드 에스터의 조성으로부터 헥사 및 옥타펩타이드의 비율은 감소하였고 디펩타이드와 테트라펩타이드의 글리세릴 에스터가 반응 생성물을 주로 구성되어 있으며 특히 AG-OGl의 조성이 증가하였음을 확인하였다. 글리세릴 펩타이드를 양이온교환수지 칼럼을 사용하여 미반응된 펩타이드와 분리하여 17.8%의 수율로 수득하였고, 이 시료를 FT-IR로 분석하여 펩타이드의 C말단의 $COO^-$의 감소를 확인하여 글리세릴 펩타이드임을 재확인하였다.

• The Plant Pathology Journal
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• 제24권4호
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• pp.417-422
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• 2008

Apicidin is a cyclic tetrapeptide produced by some Fusarium species and is known to inhibit Apicomplexan histone deacetylase. The goals of this study were to determine species identity of Fusarium isolate KCTC16676, an apicidin producer, to improve a method for apicidin extraction, and to test phytotoxicity of apicidin and its analogs. We compared sequences of the translation elongation factor 1-alpha (TEF) gene in KCTC16676 with those from isolates representing diverse Fusarium species, which showed that KCTC16676 belongs to the F. semitectum-F. equiseti species complex. To enhance apicidin production, after culturing isolate KCTC16676 on a wheat medium for 3 weeks at $25^{\circ}C$, the culture was extracted with chloroform. Apicidins were purified through a reverse phase $C_{18}$ silica gel column, resulting in 5 g of apicidin, 200 mg of apicidin A, and 300 mg of apicidin $D_2$ from 4 kg of wheat cultures; this represents a significant yield improvement from a previous method, offers more materials to study the modes of its action, and facilitates the elucidation of the apicidin biosynthesis pathway. Apicidin and apicidin $D_2$ showed phytotoxicity on both seedlings and 2-week-old plants of diverse species, and weeds were more sensitive to apicidins than vegetables

• KSBB Journal
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• 제26권6호
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• pp.483-490
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• 2011

Bioactive peptides (BAP) showed excellent cosmetic activity than bio-materials such as caffeic acid (CA), gallic acid (GA), and nicotinic acid (NA). Caffeoyl tripeptide-1 (CT-1) is a BAP that is stabilized with Gly-His-Lys (GHK) tripeptide and CA by using Fmoc solid phase peptide synthesis. Digalloyl tetrapeptide-19 (DT-19) is stabilized by combining Lys-Glu-Cys-Gly with GA and nicotinoyl tripeptide-1 (NT-1) is synthesized by GHK and NA. According to experiments, CT-1 has an excellent anti-oxidant function even with a very small amount of 10 ppm CT-1. DT-19's tyrosinase inhibition activity has the better effect of about 28.57% in 0.01% and 33.33% in 0.005% of concentration and about 7.89% in 0.001% concentration than vitamin-C. In addition, NT-1 is safer than the NA. Almost BAPs like pal-KTTKS, acetyl hexapeptide, and copper tripeptide-1 have the anti-wrinkle effect while DT-19 and NT-1 are applicable for potential BAPs focused on the whitening effect. The three kinds of BAPs like CT-1, DT-19, and NT-1 consisting of amino acids are safe to the skin, and have more excellent stability than bio-materials which are found to be unstable and cause skin irritation. Due to the high biological activity of BAP in the field of skin care, its utilization will increase constantly.

• 방사선산업학회지
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• 제2권3호
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• pp.97-104
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• 2008

Cinnamate-4-hydroxylase (C4H) is a key enzyme of phenylpropanoid pathway, which leads a variety of secondary metabolites to participate in differentiation and protection of plant against environmental stresses. In this study, we isolated a full-length cDNA of the C4H gene from a black raspberry (Rubus occidentalis L.), using a reverse transcriptase-PCR and rapid amplification of the cDNA ends (RACE)-PCR. The full-length cDNA of the RocC4H gene contained a 1,515 bp open reading frame (ORF) encoding a 504 amino acid protein with a calculated molecular weight of about 57.9 kDa and an isoelectric point (pI) value of 9.1. The genomic DNA analysis revealed that RocC4H gene had three exons and two introns. By multiple sequence alignment, RocC4H protein was highly homologous with other plant C4Hs, and the cytochrome P450-featured motifs, such as the heme-binding domain, the T-containing binding pocket motif (AAIETT), the ERR triad, and the tetrapeptide (PPGP) hinge motif, were highly conserved. Southern blot analysis revealed that RocC4H is a single copy gene in R. occidentalis.

• Biomolecules & Therapeutics
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• 제27권5호
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• pp.474-483
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• 2019

Vascular endothelial growth factor (VEGF) plays a pivotal role in pathologic ocular neovascularization and vascular leakage via activation of VEGF receptor 2 (VEGFR2). This study was undertaken to evaluate the therapeutic mechanisms and effects of the tetrapeptide Arg-Leu-Tyr-Glu (RLYE), a VEGFR2 inhibitor, in the development of vascular permeability and choroidal neovascularization (CNV). In cultured human retinal microvascular endothelial cells (HRMECs), treatment with RLYE blocked VEGF-A-induced phosphorylation of VEGFR2, Akt, ERK, and endothelial nitric oxide synthase (eNOS), leading to suppression of VEGF-A-mediated hyper-production of NO. Treatment with RLYE also inhibited VEGF-A-stimulated angiogenic processes (migration, proliferation, and tube formation) and the hyperpermeability of HRMECs, in addition to attenuating VEGF-A-induced angiogenesis and vascular permeability in mice. The anti-vascular permeability activity of RLYE was correlated with enhanced stability and positioning of the junction proteins VE-cadherin, ${\beta}$-catenin, claudin-5, and ZO-1, critical components of the cortical actin ring structure and retinal endothelial barrier, at the boundary between HRMECs stimulated with VEGF-A. Furthermore, intravitreally injected RLYE bound to retinal microvascular endothelium and inhibited laser-induced CNV in mice. These findings suggest that RLYE has potential as a therapeutic drug for the treatment of CNV by preventing VEGFR2-mediated vascular leakage and angiogenesis.

• 대한화장품학회지
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• 제49권3호
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• pp.193-201
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• 2023

H₂O₂는 세포에서 산화 스트레스를 유발하여 세포 성장, 증식, 노화 및 사멸에 영향을 미치는 일종의 활성산소종(ROS)이다. 본 연구의 목적은 H₂O₂의 세포 독성을 완화시키는 활성 펩타이드를 찾는 것이다. 잠재적인 활성 펩타이드의 서열을 예측하기 위해서 위치 스캐닝 합성 테트라펩타이드 조합 라이브러리를 탐색하였다. H₂O₂로 유도된 인간 각질형성세포(HaCaT 세포)의 사멸에 대한 펩타이드 풀들의 완화 효과를 비교한 결과, 다양한 활성 펩타이드의 시퀀스가 예측되었다. 특히 시스테인(C) 잔기를 함유한 펩타이드가 활성이 있을 것으로 예측되었다. 후속 실험에서 C-NH₂, CC-NH₂, CCC-NH₂, CCCC-NH₂ 등의 길이가 다른 시스테인 함유 펩타이드들의 세포 독성과 활성을 조사하였다. C-NH₂ 및 CC-NH₂는 1.0 mM 이하에서 유의한 세포 독성이 없었지만 CCC-NH₂ 및 CCCC-NH₂는 상대적으로 강한 세포 독성을 보였다. C-NH₂와 CC-NH₂는 H₂O₂로 유도된 세포독성을 완화시켰다. CC-NH₂는 C-NH₂, C, N-아세틸 시스테인(NAC) 및 글루타티온(GSH)보다 세포 보호 효과가 높았다. 유세포 분석법으로 세포 내 ROS를 측정하였을 때, H₂O₂는 ROS의 생성을 증가시켰다.H₂O₂ 노출조건에서 CC-NH₂는 C-NH₂보다는 더 효과적으로 ROS 생성을 억제하였고, C, NAC, GSH 만큼 효과적이었다. 본 연구의 결과는 다양한 시스테인 함유 펩타이드 중 특히 CC-NH₂가 H₂O₂로 유도된 세포 독성과 ROS 생성을 안전하고 효과적으로 완화시키는 항산화 특성이 있음을 시사한다.