• The Korean Journal of Physiology and Pharmacology
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• 제15권3호
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• pp.143-147
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• 2011

Defensins, cysteine-rich cationic polypeptides released from neutrophils, are known to have powerful antimicrobial properties. In this study, we sacrificed 30 rats to investigate the effects of ${\alpha}$-defensin 1 on detrusor muscle contractions in isolated rat bladder. From the experiments we found relaxing effects of ${\alpha}$-defensin 1 on the contractions induced by phenylephrine (PE) but not by bethanechol (BCh) in the detrusor smooth muscles. To determine the mechanisms of the effects of ${\alpha}$-defensin 1, the changes of effects on PE-induced contraction by ${\alpha}$-defensin 1 pretreatment were observed after pretreatment of Rho kinase inhibitor (Y-27632), protein kinase C (PKC) inhibitor (Calphostin C), potent activator of PKC (PDBu; phorbol 12,13-dibutyrate), and NF-${\kappa}B$ inhibitors (PDTC; pyrrolidinedithiocarbamate and sulfasalazine). The contractile responses of PE ($10^{-9}{\sim}10^{-4}$ M) were significantly decreased in some concentrations of ${\alpha}$-defensin 1 ($5{\times}10^{-9}$ and $5{\times}10^{-8}$ M). When strips were pretreated with NF-kB inhibitors (PDTC and sulfasalazine; $10^{-7}{\sim}10^{-6}$ M), the relaxing responses by ${\alpha}$-defensin 1 pretreatment were disappeared. The present study demonstrated that ${\alpha}$-defensin 1 has relaxing effects on the contractions of rat detrusor muscles, through NF-${\kappa}B$ pathway. Further studies in vivo are required to clarify whether ${\alpha}$-defensin 1 might be clinically related with bladder dysfunction by inflammation process.

• 한국응용곤충학회지
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• 제27권4호
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• pp.211-218
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• 1988

담배거세미나방(Spodoptera litura: SI), 누에(Bombyx mori: Bm) 및 흰불나방(Hyphantria cunea : Hc)으로부터 분리된 핵다각체병 바이러스(nuclear polyhedrosis virus : NPV)의 다각체의 단백질의 특성과 그에 대한 alkaline protease의 영향을 SDS-PAGE 및 혈정학적 방법으로 분석했다. Alkaline protease를 부활화시킨 후 다가체 단백질을 SDS-PAGE한 결과, BmNPV의 경우 30kD, SINPV와 HcNPV는 31kD의 단일 major band 및 이것들의 종합체(polymer)인 57kD와 66kD의 minor band들이 관찰되었다. Alkaline protease의 부활화를 성략한 SI NPV 다각체를 알칼리 용액으로 시간 차이를 두고 처리한 후 SDS-PAGE한 결고, 알칼리 처리시간이 경과함에 따라서 alkaline protease활성에 의해 다각체 단백질이 일정한 pattern으로 저분자화됨이 뚜렷하였다. SINPV 와 BmNPV 다각체 단백질의 항체를 제조하여 SINPV, BmNPV및 HcNPV 다각체 단백질간의 혈정학적 상동성을 이중형역광산법과 Western blot으로 비교한 결과는 3종 모두에서 공통 antigenic determinants의 존재가 인정되었으며 major 다각체 단백질의 종합체 형성과 alkaline protease에 의한 분해를 확인했다.

• 한국응용곤충학회지
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• 제30권3호
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• pp.180-186
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• 1991

배추흰나비 P. rapae와 P. brassicae GV에 대한 봉입체 및 바이러스단백질의 물리화학적 성상을 비교 분석한 결과 전기영동법에 의한 봉입체단백질은 그 분자량이 P. rapae GV는 30kd, P. brassicae GV는 31 kb의 단일단백질이었으며, trypsin, chymotrypsin, papain 및 staphylococcus aureus V8 protease에 의한 peptide mapping에서는 두 바이러스 간에 큰 차이가 없었다. 바이러스입자단백지르이 전기영동법에 의한 분석결과, 두 바이러스 간에 큰 차이가 없었다. 바이러스입자단백질의 전기영동법에 의한 분석결과, 두 바이러스에서 각각 42개의 band가 나타났으며 고분자량 부분에서 P. rapae GV의 경우는 115, 110, 105 및 103 kd의 4개 band인데 반해 P. brassicae GV는 107 kd의 한 band만 관찰되었다. 그리고 봉입체단백질의 아미노산 분석비교에서 두 바이러스 간의 큰 차이는 없었으며 일반적으로 곤충에 많이 존재하는 산성아미노산인 aspartic acid와 glutamic acid의 함량이 많았고 염기성 아미노산에서 lysine의 함량이 특히 많았으며 봉입체단백질의 면역학적인 시험결과에서는 P. rapae GV 항혈청에 대해 두 바이러스는 공통 침강선을 형성하였다.

• 한국응용곤충학회지
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• 제30권3호
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• pp.219-226
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• 1991

담배나방 유충에서 분리한 세포질다각체병 바이러스의 형태, 다각체 단백질 및 핵산의 전기영동상과 바이러스의 병원성을 조사하여 본 바이러스를 이용한 담배나방의 생물적 방제 이용성을 검토하고자 본 실험을 수행하였다. 다각체의 형태는 외관상 6각형으로 0.5~3.7 ${\mu}m$ 크기이고 바이러스 입자는 정 20면체로 55nm였다. SDS-PAGE에 의한 다각체 단백질은 단일 롤리？타이드인 24.3 Kd와 5개의 작은 구성분으로 이루어졌다. 바이러스입자는 7개의 폴리？타이드로 구성되어 있으며 분자량은 28.0~133.6 Kd였다. 바이러스 게놈은 10개의 조각으로 된 총 분자량 18.08 Md인 이본쇄 RNA로 각 조각의 분자량 범위는 0.65~2.79 Md이였다. 3령 유충에 대한 담배나방 세포질 다각체병바이러스의 $LC_{50}$은 $2.895{\times}10^5PIBs/ml$이었으며 $5.0{\times}10^{6}PIBs/ml$의 농도에서 $LT_{50}$에서 16.4일이었다.

• BMB Reports
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• 제35권2호
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• pp.186-193
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• 2002

The acid-bible subunit (ALS) associates with the insulinlike growth factor (IGF)-I or II, and the IGF binding protein-3 (IGFBP-3) in order to form a 150-kD complex in the circulation. This complex may regulate the serum IGFs by restricting them in the vascular system and promoting their endocrine actions. Little is known about how ALS binds to IGFBP3, which connects the IGFs to ALS. Xenopus oocyte was utilized to study the function of ALS in assembling IGFs into the ternary complexes. Xenopus oocyte was shown to correctly translate in vitro transcribed mRNAs of ALS and IGFBP3. IGFBP3 and ALS mRNAs were injected in a mixture, and their products were immunoprecipitated by antisera against ALS and IGFBP3. Contrary to traditional reports that ALS interacts only with IGF-bound IGFBP3, this study shows that ALS is capable of forming a binary complex with IGFBP3 in the absence of IGF When cross-linked by disuccinimidyl suberate, the band that represents the ALS-IGFBP3 complex was evident on the PAGE. IGFBP3 movement was monitored according to the distribution between the hemispheres. Following a localized translation in the vegetal hemisphere, IGFBP3 remained in the vegetal half in the presence of ALS. However, the mutant IGFBP3 freely diffused into the animal half, despite the presence of ALS, which is different from the wild type IGFBP3. This study, therefore, suggests that ALS may play an important role in sequestering IGFBP3 polypeptides via the intermolecular aggregation. Studies using this heterologous model will lead to a better understanding of the IGFBP3 and ALS that assemble into the ternary structure and circulate the IGF system.

• BMB Reports
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• 제41권11호
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• pp.814-819
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• 2008

Interleukin-32 (IL-32) induces a variety of proinflammatory cytokines and chemokines. The IL-32 transcript was reported originally in activated T cells; subsequently, it was demonstrated to be abundantly expressed in epithelial and endothelial cells upon stimulation with inflammatory cytokines. IL-32 is regulated robustly by other major proinflammatory cytokines, thereby suggesting that IL-32 is crucial to inflammation and immune responses. Recently, an IL-32$\alpha$-affinity column was employed in order to isolate an IL-32 binding protein, neutrophil proteinase 3 (PR3). Proteinase 3 processes a variety of inflammatory cytokines, including TNF$\alpha$, IL-$1{\beta}$, IL-8, and IL-32, thereby enhancing their biological activities. In the current study, we designed four PR3-cleaved IL-32 separate domains, identified by potential PR3 cleavage sites in the IL-32$\alpha$ and $\gamma$ polypeptides. The separate domains of the IL-32 isoforms $\alpha$ and $\gamma$ were more active than the intrinsic $\alpha$ and $\gamma$ isoforms. Interestingly, the N-terminal IL-32 isoform $\gamma$ separate domain evidenced the highest levels of biological activity among the IL-32 separate domains.

• Journal of Microbiology
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• 제38권2호
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• pp.80-87
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• 2000

The nucleotide sequence of the luxC gene coding for lux-specific fatty acyl-CoA reductase and the upstream DNA (325bp)of the structural gene from bioluminescent bacterium, Photobacterium phosphoreum, has been deternubed. An open reading frame extending for more than 20 codons in 325 bp DNA upstream of luxC was not present in both directions. The lux gene can be translated into a polypeptide of 54 kDa and the amino acid sequences of lux specific reductases of P. phosphoreum shares 80, 65, 58, and 62% identity with those of the Photobacterium leiognathi, Vibrio fischeri, Vibrio harveyi, and Xehnorhabdus luminescenens reductases, respectively. Analyses of codon usage, showing that a high frequency (2.3%) of the isoleucine codon, AUA, in the luxC gene compared to that found in Escherichia coli genes (0.2%) and its absence in the luxA and B genes, suggested that the AUA codon may play a modulator role in the expression of lux gene in E. coli. The structural genes (luxC, D, A, B, E) of the P. phosphoreum coding for luciferase (${\alpha}$,${\beta}$) and fatty acid reductase (r, s, t) polypeptides can be expressed exclusively in E. coli under the T7 phage RNA polymerase/promoter system and identificationof the [35S]methionine labelled polypeptide products. The degree of expression of lux genes in analyses of codon usage. High expression of the luxC gene could only be accomplished in a mutant E. coli 43R. Even in crude extracts, the acylated acyl-CoA reductase intermediate as well as acyl-CoA reductrase activities could be readily detected.

• 미생물학회지
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• 제50권4호
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• pp.327-333
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• 2014

산성 배지에서 성장하며 mannanase를 생산하는 균으로 분리된 Bacillus sp. YB-1401과 B. amyloliquefaciens YB-1402로부터 mannanase 유전자를 각각 클로닝하고 그 염기서열을 분석한 결과 2개 유전자는 동일하게 360 아미노산으로 구성된 단백질을 코드하며 1,080 뉴클레오티드로 구성되어 있다. 아미노산 잔기 배열의 유사성을 분석한 결과 이들 mannanases는 서로 4개의 잔기가 다르며 glycosyl hydrolase family 26에 속하는 mannanase와 상동성이 높았다. 재조합 대장균이 생산하는 YB-1402 유래의 His-tagged mannanase (HtMAN1402)가 YB-1401 유래의 His-tagged mannanase (HtMAN1401)에 비해 열안정성과 산성 pH에서 안정성이 높았다. 특히 HtMAN1402는 pH 3.0에서 4시간 방치 후에도 약 50% 이상의 잔존활성을 보여 사료첨가용 효소로 사용되기에 적합한 특성을 보였다. 또한 이들 효소는 $Ca^{2+}$ 이온에 의해 열안정성이 증가하는 것으로 확인되었다.

• 대한예방한의학회지
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• 제14권1호
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• pp.97-110
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• 2010

The wound healing process can be categorized as follows : inflammation, fibroplasia, neovascularization, collagen deposition, epithelialization, and wound contraction. During the healing process, various growth factors are secreted to accelerate wound healing. Previous studies have demonstrated that endogenous growth factors, such as vascular endothelial growth factor(VEGF) are the important regulatory polypeptides for coordinating the healing process. They are released from macrophages, fibroblasts, and keratinocytes at the site of injury and participate in the regulation of reepithelization, granulation tissue formation, collagen synthesis and neovascularization. Onchung-Um has been used clinically to treat various skin diseases. In addition, Onchung-Um has been also used for congestive inflammations. In the present study, we evaluated the effects of Onchung-Um on wound healing process and wound size reduction in rats. Full-thickness skin wounds ($15mm\;{\times}\;15mm$) were created on the back of rats. Rats were then divided into 2 groups : The Onchung-Um treated group that was orally administered with a dose of 193.9mg/100g of Onchung-Um extract per day for 15 days and Control group without Onchung-Um administration. Moreover, the histological changes and VEGF immunoexpressions of two groups were estimated. In results, wound closures were significantly accelerated by oral administration of Onchung-Um extract. Furthermore, in Onchung-Um treated group, there were significant increases in fibroblast migration, epithelialization compared with the Control group. VEGF expressions were also increased in Onchung-Um treated group. This study has therefore demonstrated the Onchung-Um can significantly improve the quality of wound healing and scar formation and the oral administration of Onchung-Um extract may increase early tissue angiogenesis in the incisional wound of an experimental animal model.

• Journal of Animal Science and Technology
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• 제59권9호
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• pp.20.1-20.9
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• 2017

Background: Ribonuclease (RNase) is one of the few toxic proteins that are present constantly in snake venoms of all types. However, to date this RNase is still poorly studied in comparison not only with other toxic proteins of snake venom, but also with the enzymes of RNase group. The objective of this paper was to investigate some properties of RNase from venom of Vietnam cobra Naja atra. Methods: Kinetic methods and gel filtration chromatography were used to investigate RNase from venom of Vietnam cobra. Results: RNase from venom of Vietnam cobra Naja atra has some characteristic properties. This RNase is a thermostable enzyme and has high conformational stability. This is the only acidic enzyme of the RNase A superfamily exhibiting a high catalytic activity in the pH range of 1-4, with $pH_{opt}=2.58{\pm}0.35$. Its activity is considerably reduced with increasing ionic strength of reaction mixture. Venom proteins are separated by gel filtration into four peaks with ribonucleolytic activity, which is abnormally distributed among the isoforms: only a small part of the RNase activity is present in fractions of proteins with molecular weights of 12-15 kDa and more than 30 kDa, but most of the enzyme activity is detected in fractions of polypeptides, having molecular weights of less than 9 kDa, that is unexpected. Conclusions: RNase from the venom of Vietnam cobra is a unique member of RNase A superfamily according to its acidic optimum pH ($pH_{opt}=2.58{\pm}0.35$) and extremely low molecular weights of its major isoforms (approximately 8.95 kDa for RNase III and 5.93 kDa for RNase IV).