• Molecules and Cells
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• 제21권2호
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• pp.229-236
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• 2006

Gaegurin 4 (GGN4), a novel peptide isolated from the skin of a Korean frog, Rana rugosa, has broad spectrum antimicrobial activity. A number of amphipathic peptides closely related to GGN4 undergo a coil to helix transition with concomitant oligomerization in lipid membranes or membrane-mimicking environments. Despite intensive study of their secondary structures, the oligomeric states of the peptides before and after the transition are not well understood. To clarify the structural basis of its antibiotic action, we used analytical ultracentrifugation to define the aggregation state of GGN4 in water, ethyl alcohol, and 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP). The maximum size of GGN4 in 15% HFIP corresponded to a decamer, whereas it was monomeric in buffer. The oligomeric transition is accompanied by a cooperative 9 nm blue-shift of maximum fluorescence emission and a large secondary structure change from an almost random coil to an ${\alpha}$-helical structure. GGN4 induces pores in lipid membranes and, using electrophysiological methods, we estimated the diameter of the pores to be exceed $7.3{\AA}$, which suggests that the minimal oligomer structure responsible is a pentamer.

• 농업과학연구
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• 제42권1호
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• pp.29-35
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• 2015

This study was performed to analyze the distribution and antimicrobial resistance of pathogenic microorganisms isolated from the carcass and environments of chicken processing plant located in Gyeonggi province from October to November in 2010. Chicken slaughterhouse was visited 3 times and totally 40 samples were collected from chicken carcass before and after washing (n=14), chicken cuts (n=7), cooling water (n=8), brine (n=2), cutting knives (n=7) and working plate (n=2). Whole-chicken rinsing technique (for chicken carcasses) and swab technique (for working plate and knives) were used to analyze the distribution of pathogenic microorganisms. In addition, brine and chilling water from storage tanks were gathered using sterilized tubes and used as samples. The matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for whole cell fingerprinting in combination with a dedicated bioinformatic software tool was used to identify the isolated microorganisms. The pathogenic microorganisms, such as Bacillus cereus (n=8) and Staphylococcus aureus (n=9), were isolated form the chicken processing process (chicken carcasses of before and after chilling, chicken cuts, and working plate). The antimicrobial susceptibility of those isolated microorganisms was analyzed using 21 antimicrobial agents. In the case of B. cereus, it showed 100% of resistance to subclasses of penicillins and peptides, and it also resistant to cephalothin, a member of critically important antimicrobials (CIA), however there was no resistance (100% susceptible) to vancomycin and chloramphenicol. S. aureus showed 100% resistance to subclasses of peptides and some of penicillins (penicillin and oxacillin), however, it showed 100% susceptibility to cephalosporins (cefazolin and cephalothin). All of the tested pathogens showed multi drug resistance (MDR) more than 4 subclasses and one of B. cereus and S. aureus showed resistance to 9 subclasses. After the ban on using the antimicrobials in animal feed in July 2011, there would be some change in microbial distribution and antimicrobial resistance, and it still has a need to be analyzed.

• 생명과학회지
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• 제34권7호
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• pp.493-499
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• 2024

항균 펩타이드는 동물과 식물이 가지고 있는 고유의 항균 물질로 적은 양으로도 강한 항균활성을 나타내며 이외에도 항바이러스, 항산화 등 다양한 기능을 가지고 있다. 식물은 물과 햇빛으로 키울 수 있어 적은 비용으로 대량 생산이 가능하다. 식물의 엽록체를 형질전환 시켜 항균 펩타이드를 생산하면 단백질 발현량이 증가하고 꽃가루에 의한 유전자 이동이 일어나지 않기 때문에 생태계가 오염될 가능성이 적다. 그러나 형질전환 된 엽록체를 이용하여 재조합 단백질을 생산하면 단백질이 분해되고 용해도가 감소한다. 이를 해결하기 위해 융합 단백질 종류 중 하나인 SUMO를 발현시킬 재조합 단백질과 융합하여 제작하였다. 항균 펩타이드 stomoxyn은 침파리(stable fly)에 있는 항균물질이다. Stomoxyn은 α-helix 구조이고 양친매성이어서 박테리아 세포막에 부착된 후 세포막을 용해시킨다. 본 연구에서는 stomoxyn을 식물 엽록체와 대장균에서 발현시키기 위한 형질전환 벡터를 제작하였고, 이 벡터를 이용하여 대장균에서 stomoxyn의 발현을 확인하였다. 대장균에서 발현된 stomoxyn을 nickel column과 SUMOase를 처리하여 정제한 후 agar diffusion assay를 이용하여 항균 활성을 확인하였다. 또한 식물 엽록체에서 벡터의 삽입을 확인하기 위해 EGFP 유전자를 사용하여 확인하였다.

• BMB Reports
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• 제48권8호
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• pp.479-484
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• 2015

In this study, we report that one of the antimicrobial peptides scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates actin polymerization and the subsequent chemotactic migration of macrophages through the activation of ERK and protein kinase B (Akt) activity. The scolopendrasin VII-induced chemotactic migration of macrophages is inhibited by the formyl peptide receptor 1 (FPR1) antagonist cyclosporine H. We also found that scolopendrasin VII stimulate the chemotactic migration of FPR1-transfected RBL-2H3 cells, but not that of vector-transfected cells; moreover, scolopendrasin VII directly binds to FPR1. Our findings therefore suggest that the antimicrobial peptide scolopendrasin VII, derived from Scolopendra subspinipes mutilans, stimulates macrophages, resulting in chemotactic migration via FPR1 signaling, and the peptide can be useful in the study of FPR1-related biological responses. [BMB Reports 2015; 48(8): 479-484]

• Bulletin of the Korean Chemical Society
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• 제30권8호
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• pp.1839-1844
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• 2009

A 20-residue hybrid peptide CA(1-8)-MA(1-12) (CAMA) incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA) has high antimicrobial activity without toxicity. To investigate the effects of the total positive charges of CAMA on the antibacterial activity and toxicity, a hybrid peptide analogue (CAMA-syn) was designed with substitutions of $Ile^{10}\;and\;Ser^{16}$ with Lys. According to CD spectra, structure of CAMA-syn with increase of cationicity was very similar to that of CAMA in DPC micelle. CAMA-syn showed antimicrobial activity similar with CAMA while CAMA-syn has no hemolytic activity and much lower cytotoxicity against RAW 264.7 macrophage cells than CAMA. Also, CAMA and CAMA-syn significantly inhibited NO production by LPSstimulated RAW264.7 macrophage at 10.0∼20.0 $\mu$M. CAMA-syn displayed salt resistance on antimicrobial activity against Escherichia coli at the physiological concentrations of $CaCl_2\;and\;MgCl_2$. The combination studies of peptides and antibiotics showed that CAMA-syn has synergistic effects with synthetic compound and flavonoid against Enterococcus faecalis and VREF. CAMA-syn can be a good candidate for the development of new antibiotics with potent antibacterial and synergistic activity but without cytotoxicity.

• BMB Reports
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• 제32권1호
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• pp.12-19
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• 1999

Gaegurin 4 (GGN4), a broad-spectrum antibiotic, is a 37-amino acid peptide isolated from the Korean frog, Rana rugosa. In this study, we have chemically synthesized and purified GGN4 analogues where the C-terminal portion is truncated and/or substituted with tryptophan. These peptides show significantly different biological activities depending on the location of tryptophan and the number of amino acids truncated from the C-terminal end. While deletion of 9 amino acids from the C-terminal seems to be marginally tolerable in maintaining the antimicrobial activity, further deletion of up to 14 amino acid residues decreases the potency by more than 60-fold towards Gram-positive, and 10-fold towards Gram-negative, bacteria. Surprisingly, the reduced activity of the shorter peptide can be completely restored by a single substitution of aspartic acid 16 to tryptophan 16 (D16W). Also, the truncation seems to decrease the specificity of antibiotic activity more towards Gram-positive than towards Gram-negative bacteria studied. These data suggest a partial role of the C-terminal region in determining the binding specificity and the activity of peptides upon binding to their target cell membranes.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2010년도 정기총회 및 추계학술발표회
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• pp.12-12
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• 2010

Nature applied flavanoids, glucosinolates and peptides for a great variety of functions. Flavanoids and glucosinolates are rich source of antioxidants, generally present in colored fruits and vegetables. Quercetin and its glucosides areone of the best examples of flavonol used in daily diet. Whereas peptides can act as antimicrobial, chemical messenger, neurotransmitter etc that regulating various life processes. Aspartame, a dipeptide is used as artificial sweetener and oxytocin for medical purposes, gained importance in everyday life. So, flavanoids, glucosinolates, peptides and their derivatives continue to hold the attention of synthetic chemists, agriculturists and biologists. Apart from a variety of naturally occurring bioactive metabolites, we are aiming to extract, separate and synthesize new analogs of promising natural drug candidates.

• Bulletin of the Korean Chemical Society
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• 제35권9호
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• pp.2809-2812
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• 2014

The 43-residue defensin-like peptide coprisin, which is isolated from dung bettle, Copris tripartitus, is a potent antimicrobial peptide. In our previous work, we determined the tertiary structure of coprisin and found that alpha helical region of coprisin from residue 19 to residue 30 is important for its antimicrobial activities. Here, we designed cop12mer and cop9mer analogs of coprisin based on the tertiary structure of coprisin. To investigate the relationship between hydrophobicity and antimicrobial activities and develop the potent peptide antibiotics, we designed cop9mer-1 with substitution of $His^2$ with Trp in cop9mer. The results showed that cop9mer-1 has higher toxicities as well as improved antimicrobial activities compared to cop9mer. In order to reduce the toxicity of cop9mer-1, we designed cop9mer-2 and cop9mer-3 with substitution of $Cys^3$ with Lys or Ser. Substitution of $Cys^3$ with these hydrophilic amino acids results in lower cytotoxicities compared to cop9mer-1. Cop9mer-2 with substitution of $Cys^3$ with Lys in Cop9mer-1 showed high antibacterial activities against drug resistant bacteria without cytotoxicity. Antibiotic action of cop9mer-1 analog appears to involve permeabilization of the bacterial cell membrane while cop9mer-2 and cop9mer-3 may have different mechanism of action. These results imply that that optimum balance in hydrophobicity and hydrophilicity in these 9-meric peptides plays key roles in their antimicrobial activities as well as cytotoxicities.

• 대한화학회지
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• 제54권2호
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• pp.183-191
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• 2010

1-Palmitoyl-$d_{31}$-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC_$d_{31}$)로 이루어진 지질 이중막에 항균성 펩타이드 magainin 2와 aurein 3.3이 작용했을 때 일어나는 상변화를 중수소 고체 핵자기 공명 스펙트럼을 관측하여 탐구하였다. 측정된 중수소 고체 핵자기 공명 스펙트럼의 선모양을 이론적 모델 구조를 통하여 모사함으로써, 펩타이드-지질 혼합상태의 기하학적 구조상수 및 동력학적 표면 확산 계수 등을 구하였다. 펩타이드의 혼합 이후 5일 이내의 짧은 작용시간에는 지질 이중막의 정렬이 파괴되는 현상이 나타났으나, 100일 이후에는 magainin 2에 의해서는 타원형 원환체 기공 (elliptic toroidal pore)이, aurein 3.3에 의해서는 6각형 단면 막대구조 상(hexagonal phase)이 관측되었다. 지질 이중막 표면의 전하밀도가 항균성 펩타이드의 작용에 미치는 효과를 보기 위해 산성 지질인 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG)를 함유한 POPC_$d_{31}$/POPG 지질 이중막에 대해서도 동일한 실험을 수행하였다. 동일한 ${\alpha}$-나선형 구조를 가지는 두 펩타이드가 두 종류의 지질 이중막에 작용하는 과정의 차이를 확인할 수 있었다.

• 생명과학회지
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• 제25권7호
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• pp.833-838
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• 2015

다제약제 내성을 가지는 슈퍼 박테리아, 곰팡이, 바이러스, 기생충 감염은 주요한 건강위협인자들이다. 하지만, 건강위협 상황을 극복하기위해 현재 약제의 대안들중 항생펩타이드를 들 수 있다. 항생펩타이드는 자연계 다양한 종에서 생산된다. 항생펩타이드는 작은 단백질로 어류, 양서류, 파충류, 식물 그리고 동물의 감염으로부터 다세포생명체를 보호하는 선천성 면역에 관여하고 있다. 1980년대 이후로 매년 항생펩타이드의 수가 증가하고 있다. 박테리아, 원생생물, 곰팡이, 식물, 동물로부터 동정된 2,000가지 이상의 항생펩타이드가 항생펩타이드 데이터베이스에 등록되어 있다. 이러한 항생펩타이드의 대부분은 11-50개의 아미노산으로 구성되어 있고 하전상태는 0에서 +7까지이며 소수성은 31-70%를 차지하고 있다. 본 보고는 항생펩타이드를 생물학적 원천, 생물학적 기능, 펩타이드 성질, 공유결합패턴, 3차구조등에 의해 분류하였다. 항생펩타이드의 기능은 항균작용외에 세포주화성과 같은 세포생물학적 활성에도 기능성을 가지고 있다. 더욱이 림프절 스트로마로부터 기원한 fibroblastic reticular cell (FRC)에 염증상황 유도시 항생펩타이드가 발현되는 것을 확인하였다. FRC로부터 유도된 항생펩타이드는 whey acidic protein (WAP) 도메인을 포함하고 있었다. 이것은 단백질 도메인에 의해서도 항생펩타이드를 분류 할 수 있다는 것을 제시한다.