• International Journal of Industrial Entomology and Biomaterials
• /
• 제37권2호
• /
• pp.49-54
• /
• 2018

To artificially enhance antimicrobial peptide expression in Bombyx mori, we constructed genetically engineered silkworms overexpressing Rel family transcription factor. The truncated BmRelish1 (BmRelish1t) gene contained a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acid (AHAA)-rich region, and death domain (DD), but no ankyrin-repeat (ANK) domain. The BmRelish1t gene was controlled by B. mori cytoplasmic actin 3 promoter in the PiggyBac transposon vector. Chromosome analysis of G1 generations of a transgenic silkworm with EGFP expression confirmed stable insertion of BmRelish1t. BmRelish1t gene overexpression in transgenic silkworms resulted in higher mRNA expression levels of B. mori antimicrobial peptides such as lebocin(~20.5-fold), moricin(~8.7-fold), and nuecin(~17.4-fold) than those in normal silkworms.

• 생명과학회지
• /
• 제28권11호
• /
• pp.1301-1315
• /
• 2018

참담치(Mytilus coruscus)의 혈구 유래의 항균 펩타이드를 역상 column들을 사용한 reversed-phase high-performance liquid chromatography (RP-HPLC)로 분리 및 정제하였다. 정제된 펩타이드는 matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS)를 통해 분자량이 4041.866 Da으로 밝혀졌으며 Edman degradation법을 통해 25개의 N-말단 서열을 확보하였다. 이는 참담치의 mytilin B precursor와 100%, mytilin 8 precursor, mytilin 4 precursor와 96% 일치하였다. 또한 103개의 아미노산 서열을 코딩하고 있는 312 bp의 open-reading frame (ORF)을 밝혔으며 이는 참담치의 mytilin B precursor와 100% 일치하였다. 밝혀진 분자량과 아미노산 서열을 바탕으로 C-말단 alanine 잔기의 유무에 따라 2개의 펩타이드를 합성하였으며 이는 mytilin B1과 B2라고 명명하였다. 이들은 그람 양성 균주 Bacillus cereus, Streptococcus parauberis [minimal effective concentrations, MECs $41.6-89.7{\mu}g/ml$], 그람 음성 균주 Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, Vibrio ichthyoenteri [MECs $7.4-39.5{\mu}g/ml$] 그리고 진균류인 Candida albicans [MECs $26.0-31.8{\mu}g/ml$]에 항균활성을 나타냈다. 본 연구 결과, 참담치 혈구 유래 mytilin B1과 mytilin B2는 넓은 항균 스펙트럼을 가지고 열과 염분에 대한 안정성이 높으며 용혈현상과 세포독성은 나타나지 않았다. 이러한 특성은 기능성 사료첨가제 및 항생제 대체제로써 충분히 안정적인 역할을 할 뿐만 아니라 추후 mytilin의 구조적 중요성과 참담치의 면역학적 측면에서 다양한 자료를 제시할 것으로 사료된다.

• Bulletin of the Korean Chemical Society
• /
• 제29권2호
• /
• pp.323-327
• /
• 2008

A focused library of pyrazole-based compounds was constructed towards novel transcription factor inhibitors. Complementary hydrogen bonding interaction with b-sheet peptide structures was the basis for the design of 5-amino-3-pyrazole carboxamide scaffold. From the preliminary inhibition assay against several transcription factors, compounds 7e and 8g were identified as novel lead compounds against HIF-1a and NF-AT transcription factors, respectively.

• 한국결정학회:학술대회논문집
• /
• 한국결정학회 2002년도 정기총회 및 추계학술연구발표회
• /
• pp.4-4
• /
• 2002

PDZ domains bind to short segments within target proteins in a sequence-specific fashion. GRIP/ABP family proteins contain six to seven PDZ domains and interact via its sixth PDZ domain (class Ⅱ) with the C-termini of various proteins, including liprin-α. In addition the PDZ456 domain mediates the formation of homo- and heteromultimers of GRIP proteins. To better understand the structural basis of peptide recognition by a class Ⅱ PDZ domain and DZ-mediated multimerization, we determined the crystal structures of the GRIPI PDZ6 domain, alone and in complex with a synthetic C-terminal octapeptide of human liprin-α, at resolutions of 1.5 Å and 1.8 Å, respectively. Remarkably, unlike other class Ⅱ PDZ domains, Ile736 at αB5 rather than conserved Leu732 at αB1 makes a direct hydrophobic contact with the side chain of the Tyr at the -2 position of the ligand. Moreover, the peptide-bound structure of PDZ6 shows a slight reorientation of helix αB, indicating that the second hydrophobic pocket undergoes a conformational adaptation to accommodate the bulkiness of the Tyr's side chain, and forms an antiparallel dimer through an interface located at a site distal to the peptide-binding groove. This configuration may enable formation of GRIP multimers and efficient clustering of GRIP-binding proteins.

• Fisheries and Aquatic Sciences
• /
• 제14권3호
• /
• pp.174-178
• /
• 2011

This study aimed to determine the degree of hydrolysis and angiotensin-I-converting enzyme (ACE)-inhibitory activity of Giant Jellyfish Nemopilema nomurai (jellyfish) hydrolysates. The degree of hydrolysis using six proteolytic enzymes (Alcalase, Flavozyme, Neutrase, papain, Protamex, and trypsin) ranged from 13.1-36.8% and the inhibitory activities from 20.46-79.58%. Using papain hydrolysate, we newly isolated and characterized ACE-inhibitory peptides with a molecular weight of 3,000-5,000 Da that originated from jellyfish collagen. The purified peptide (FII-b) was predicted to be produced from an alpha-2 fragment of the type IV collagen of jellyfish. The N-terminal sequence of FII-b was Asp-Pro-Gly-Leu-Glu-Gly-Ala-His-Gly- and showed 87% identity to the collagen type IV alpha-2 fragment of Rattus norvegicus and a predicted protein from Nematostella vectensis, indicating that the ACE-inhibitory peptide originated from the collagen hydrolysate and had an $IC_{50}$ value of 3.8 ${\mu}g$/mL. The primary structure of the fragment is now being studied; this peptide represents an interesting new type of ACE inhibitor and will provide knowledge of the potential applications of jellyfish components as therapies for hypertension.

• Journal of Plant Biotechnology
• /
• 제40권3호
• /
• pp.169-177
• /
• 2013

최근 급속도로 당뇨병 환자가 증가하면서 인슐린 시장이 크게 성장하고 있다. 또한 최근 식물체를 이용하여 의약용 단백질 생산이 경제적인 측면과 안정성 측면에서 매우 효과적임이 보고되고 있어 이를 이용한 분자농업이 주목을 받고 있다. 본 연구에서는 인슐린 단백질을 식물체에서 생산하기 위한 유전자 발현 construct를 설계하기 위한 실험으로서 식물발현용 preprominiinsulin construct를 제조하기 위한 단계적 실험을 수행하였다. 우선 proinsulin이 무세포 식물 전사/번역시스템에서 성공적으로 발현됨을 확인하였다. Prominiinsulin construct를 제조하여 대장균에서 발현시키는데 성공하였으며, 이를 트립신으로 절단한 후 인간 항인슐린 항체를 이용한 western 분석을 통하여 효과적으로 A-펩타이드와 B-펩타이드가 형성되며 이후 적절하게 접힘이 일어나고 hexamer로 조립됨을 확인하였다. 이후 식물체에서 재조합 인슐린 유전자가 발현되는지를 확인하기 위하여 RFP 결합 construct를 제조하여 담배의 현탁배양세포인 BY-2 세포에 형질전환시켜 RFP 결합 preprominiinsulin이 성공적으로 발현됨을 확인 하였다. 이러한 성공적인 연구 결과를 토대로 향후 이 construct는 RFP 단백질을 제거하여 35S 프로모터에 직접 유도되는 [N 말단 ${\rightarrow}$ tobacco E2 시그널 펩타이드 ${\rightarrow}$ B-펩타이드(1-29 AA) ${\rightarrow}$ AAK ${\rightarrow}$ A-펩타이드(1-21 AA) ${\rightarrow}$ RR ${\rightarrow}$ His6 ${\rightarrow}$ KDEL ${\rightarrow}$ C 말단] construct를 제조하여 담배 식물체에 형질전환시켜 분자농업을 통해 인간 인슐린 단백질을 생산하는데 활용하고자 한다.

• Journal of Microbiology and Biotechnology
• /
• 제22권1호
• /
• pp.156-158
• /
• 2012

A synthetic coprisin analog peptide, 9-mer dimer CopA3 (CopA3) was designed based on a defensin-like peptide, Coprisin, isolated from the bacteria-immunized dung beetle Copris tripartitus. Here, CopA3 was investigated for its antimicrobial activity and cancer cell growth inhibition. CopA3 showed antimicrobial activities against various pathogenic bacteria and yeast fungus with MIC values in 2~32 ${\mu}M$ ranges, and inhibited the cell viabilities of pancreatic and hepatocellular cancer cells, except MIA-Paca2, Hep3B, and HepG2 cells, in a dose-dependent manner. The average $IC_{50}$ values of CopA3 against pancreatic and hepatocellular cancer cells were 61.7 ${\mu}M$ and 67.8 ${\mu}M$, respectively. The results indicate that CopA3 has potential in the treatments of pancreatic and hepatocellular cancers as well as microorganism infection disease.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1995년도 제3회 추계심포지움
• /
• pp.107-113
• /
• 1995

Basement membrane laminin is a multidomain glycoprotein that interacts with itself, heparin and cells. The distal long arm plays major cell and heparin interactive roles. The long arm consists of three subunits (A, B1, B2) joined in a coiled-coil rod attached to a terminal A chain globule (G). The globule is in turn subdivided into five subdomains (Gl-5). In order to analyze the functions of this region, recombinant G domains (rG, rAiG, rG5, rGΔ2980-3028) were expressed in Sf9 insect cells using a baculovirus expression vector. A hybrid molecule (B-rAiG), consisting of recombinant A chain(rAiG) and the authentic B chains (E8-B)was assembled in vitro. The intercalation of rAiG into E8-B chains suppressed a heparin binding activity identified in subdomain Gl-2. By the peptide napping and ligand blotting, the relative affinity of each subeomain to heparin was assigned as Gl> G2= G4> G5> G3, such that G1 bound strongly and G3 not at all. The active heparin binding site of G domain in intact laminin appears to be located in G4 and proximal G5. Cell binding was examined using fibrosarcoma Cells. Cells adhered to E8, B-rAiG, rAiG and rG, did not bind on denatured substrates, poorly bound to the mixture of E8-B and rG. Anti-${\alpha}$6 and anti-${\beta}$1 integrin subunit separately blocked cell adhesion on E8 and B-rAiG, but not on rAiG. Heparin inhibited cell adhesion on rAiG, partially on B-rAiG, and not on E8. In conclusion, 1) There are active and cryptic cell and heparin binding activities in G domain. 2) Triple-helix assembly inactivates cell and heparin binding activities and restores u6131 dependent cell binding activities.

• Tuberculosis and Respiratory Diseases
• /
• 제54권3호
• /
• pp.311-319
• /
• 2003

연구배경 : B-type natriuretic peptide(BNP)는 주로 심장의 심실에서 분비되는 호르몬으로서 심부전의 진단, 중증도 및 예후와도 연관이 있다고 알려져 있다. 중환자의 경우 급성병색시 심부전이 올수 있는 것으로 알려져있고 이는 환자의 예후와 연관이 있을 가능성이 있다. 그래서 저자들은 우선 이전 심질환의 병력이 없는 중환자들에서 BNP를 측정하여 증가여부를 살펴보고, BNP와 중환자의 중등도 및 예후와의 관련성 여부를 살펴보기로 하였다. 방 법 : 2002년 6월부터 10월까지 본원 중환자실에 입원하였던 환자 중 환자의 중증도와는 관련이 없이 BNP가 증가될 수 있는 울혈성 심부전, 심방 세동, 허혈성 심질환, 신부전 등을 제외한 32명을 대상으로 하였고, 대조군으로는 일반 병동 및 외래 환자 32명을 무작위로 추출하였다. BNP는 Triage B-Type Natriuretic Peptide test를 이용하여 fluorescence immunoassay를 통해 측정하였다. APACHE II score와 APACHE III score 및 중환자실 사망 여부를 추적하였다. 결 과 : 남녀비는 16 : 16이었고, 연령은 $59.2{\pm}20.6$세 였다. BNP는 $186.7{\pm}274.1$ pg/mL으로 정상 대조군의 $19.9{\pm}21.3$ pg/mL 보다 유의하게 높았다. 중환자군에서 BNP 100 pg/mL이상은 14명으로 43.8%였다. APACHE II score는 $16.5{\pm}7.6$이었고, 11명이 사망하였다. BNP값은 APACHE II score및 사망과 유의한 상관 관계를 보였고(r=0.443, p=0.011, r=0.530, p=0.002), 생존군과 사망군 BNP값은 유의한 차이를 보였으나($83.2{\pm}55.8$ pg/mL vs. $384.1{\pm}401.7$ pg/mL ; p=0.033), $PaO_2/FiO_2$와는 상관 관계를 보이지 않았다. 결 론 : BNP는 이전 심질환병력이 없었던 중환자에서 증가되어 있었고, 사망률 및 중증도 점수와도 유의한 상관 관계가 있었다. 따라서 중환자들의 예후를 예측하는데 비침습적이고 빠른 방법으로 도움이 될 가능성이 있다.

• 생약학회지
• /
• 제16권4호
• /
• pp.233-238
• /
• 1985

A number of sedative alkaloids were isolated from Sanjoin(酸棗仁), the seeds of Zizyphus vulgaris Lamark var. spinosus Bunge (Rhamnaceae) which is an important Chinese medicinal material used to treat insomnia and sometimes to treat sleepiness. Those compounds were designated as Sanjoinine-A, B, C, D, etc. depending on the order of increasing polarity. Sanjoinine-A, $C_{31}H_{42}N_4O_4$, $mp\;249^{\circ}$, $[{\alpha}]^{27}_D-316$, Sanjoinine-B, $C_{30}H_{40}N_4O_4$, $mp\;212{\sim}4^{\circ}$, Sanjoinene, $C_{29}H_{35}N_3O_4$, $mp\;281{\sim}2^{\circ}$, $[{\alpha}]^{22}_D-272$, Sanjoinine-D, $C_{32}H_{46}N_4O_5$, $mp\;256{\sim}8^{\circ}$, $[{\alpha}]^{22}_D-53.6$, Sanjoinine-F, $C_{31}H_{42}N_4O_5$, $mp\;228{\sim}9^{\circ}$, $[{\alpha}]^{22}_D-215$, and $Sanjoinine-G_1,\;C_{31}H_{44}N_4O_5,\;mp\;236{\sim}8^{\circ},\;[{\alpha}]^{22}_D-68.6$, were found as 14-membered cyclic peptide alkaloids, $Sanjoinine-G_2,\;C_{30}H_{42}N_4O_4,\;mp\;182^{\circ},\;[{\alpha}]^{22}_D-79.2$, as being open chain peptide alkaloid, and Sanjoinine-E, $C_{19}H_{21}NO_2$, $mp\;166^{\circ}$, $[{\alpha}]^{20}_D-146.2$, N-Methylasimilobine, $C_{18}H_{19}NO_2$, $mp\;193{\sim}5^{\circ}$, $[{\alpha}]^{20}_D-204$, Sanjoinine-Ia, $C_{18}H_{19}NO_2$, $mp\;155{\sim}7^{\circ}$, $[{\alpha}]^{20}_D-140$, Sanjoinine-Ib, $C_{19}H_{21}NO_4$, $mp\;184^{\circ}$, Sanjoinine-K, $C_{16}H_{19}NO_3$, $mp\;159{\sim}61^{\circ}$, $[{\alpha}]^{20}_D+35$, Caaverine, $C_{17}H_{17}NO_2$, $mp\;204^{\circ}$, $[{\alpha}]^{20}_D-80$, and Zizyphusine, $C_{20}H_{24}NO_4$, $mp\;214{\sim}6^{\circ}$, $[{\alpha}]^{20}_D+317$ as being aporphine alkaloids. The heat treatment of the cyclic peptide alkaloids produced their isomeric products which showed enhanced sedative activity. The chemical structure of the isomeric products will be discussed.