• BMB Reports
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• 제47권10호
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• pp.552-557
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• 2014

The main purpose of this study was to investigate whether type 3 muscarinic acetylcholine receptor (M3R) dysfunction induced vascular hyperpermeability. Transwell system analysis showed that M3R inhibition by selective antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) and small interfering RNA both increased endothelial permeability. Using coimmunoprecipitation and Western blot assay, we found that M3R inhibition increased VE-cadherin and ${\beta}$-catenin tyrosine phosphorylation without affecting their expression. Using PTP1B siRNA, we found that PTP1B was required for maintaining VE-cadherin and ${\beta}$-catenin protein dephosphorylation. In addition, 4-DAMP suppressed PTP1B activity by reducing cyclic adenosine monophosphate (cAMP), but not protein kinase $C{\alpha}$ ($PKC{\alpha}$). These data indicate that M3R preserves the endothelial barrier function through a mechanism potentially maintaining PTP1B activity, keeping the adherens junction proteins (AJPs) dephosphorylation.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1634-1639
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• 2006

Hepatitis C virus (HCV)-encoded nonstructural protein 5B (NS5B) possesses RNA-dependent RNA polymerase activity, which is considered essential for viral proliferation. Thus, HCV NS5B is a good therapeutic target protein for the development of anti-HCV agents. In this study, we isolated two different kinds of nuclease-resistant RNA aptamers with 2'-fluoro pyrimidines against the HCV NS5B from a combinatorial RNA library with 40 nucleotide random sequences, using SELEX technology. The isolated RNA aptamers were observed to specifically and avidly bind the HCV NS5B with an apparent $K_d$ of 5 nM and 18 nM, respectively, in contrast with the original RNA library that hardly bound the target protein. Moreover, these aptamers could partially inhibit RNA synthesis of the HCV subgenomic replicon when transfected into Huh-7 hepatoma cell lines. These results suggest that the RNA aptamers selected in vitro could be useful not only as therapeutic agents of HCV infection but also as a powerful tool for the study of the HCV RNA-dependent RNA polymerase mechanism.

• 한국자원식물학회지
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• 제18권3호
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• pp.441-449
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• 2005

광계II(PSII)는 고등식물의 chloroplast에서 두 개의 광합성 반응중심 중의 하나이다. Chlorophyll a/b 광수확 복합체는 광계II를 위한 안테나 역할을 수행한다. 본 연구에서는 인삼의 잎조직을 제작한 cDNA library로부터 chlorophyll a/b-binding protein (Cab) 유전자를 분리하였다. 인삼 Cab유전자는 935 bp의 염기와 265개의 아미노산 잔기(pI 5.63)로 구성된 한 개의 ORF를 포함하고 있으며, 단백질의 분자량은 28.6 kDa으로 추정되었다. 인삼에서 분리한 Cab 유전자는 기존에 식물에서 보고된 유전자들과 유사성을 나타내었으며, 유사도는 $68-92\%$로 나타났다. 아미노산 서열을 비교하여 유연관계를 분석한 결과 인삼의 Cab 유전자는 비교된 P. persica (AAC34983), A.thaliana (AAD28771), G. hirsutum (CAA38025), G. max (AAL29886), V. radiata (AAF89205) 등과 동일한 그룹으로 분리되었다.

• 한국축산식품학회지
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• 제25권3호
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• pp.328-332
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• 2005

The effect of k-casein (k-CN) variant on milk production traits (milk yield, fat yield, protein yield, fat percentage and protein percentage) was estimated for 568 Holstein cows in the first lactation. The k-CN valiant were determined by PCR-RFLP (restriction fragment length polymorphism) technique at the DNA level. Single trait linear model was used for the statistical analysis of the data. Result of this study indicated that k-CN variant affected significantly milk yield (P<0.05) and protein yield (P<0.01). Animals with the BB variant produced 622kg milk more and had protein yield higher by 32kg compared with animals with the AA variant No associations between the k-CN variants and other milk production trait were found. Therefore, milk and protein yield may be improved through milk protein typing by increasing the frequencies of k-CN B variant in dairy cattle population. In cheese making, it will be also preferable to have milk with the B variant of k-CN, which gives higher yield having a better quality than the A variant milk.

• 한국가금학회지
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• 제44권2호
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• pp.67-73
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• 2017

The purpose of our study was to determine the effects of varying levels of energy, protein, and amino acids on the performances of laying hens. A total of 240 Hy-Line Brown laying hens at 36 weeks of age were used in this 4-week feeding trial. The hens were randomly allocated to five treatment diets, with eight replications of six hens in each replicate cage. The treatment diets were as follows: A- basal diet + 18% crude protein, metabolizable energy 2,800 kcal, total (methionine + cysteine) 0.65%; B- basal diet + 17% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.59%; C- basal diet + 16.5% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.59%; D- basal diet + 16.5% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.54%; and E- basal diet + 16% crude protein, metabolizable energy 2,680 kcal, total (methionine + cysteine) 0.54%. The study results revealed that the hen-day egg production of hens that were fed with low-energy diets (B, C, and D) was comparable with that of hens fed with high-energy diet A, whereas average daily feed intake in hens fed treatment diet D and E was significantly higher (P<0.05) than that in hens fed treatment diet A. Overall, the eggshell thickness was unaffected by any of the treatment diets. Egg weight was comparable among the treatment diets, except for treatment diet E. Haugh unit improved with decreasing levels of dietary energy, protein, and methionine + cysteine in the diet. We can summarize that laying hens fed with low dietary energy and low crude protein treatment diets B, C, and D had satisfactory performance compared with those fed with high-energy treatment diet A. This indicates that there is the potential to reduce feed costs by formulating diets with lower energy and low protein levels.

• 생명과학회지
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• 제21권1호
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• pp.81-88
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• 2011

누에 분말을 이용하여 B. subtilis 및 A. kawachii 균주에 의한 고체 발효를 통해 얻어진 발효 누에분말의 발효시간에 따른 이화학적 특성(단백질 함량 및 단백질 분해 패턴) 및 생리활성 효과(항산화 작용, 환원력 및 혈전용해 활성)를 평가하였다. 누에 단백질 함량과 항산화 활성은 B. subtilis 및 A. kawachii 두 균주 발효 모두 12일째에 가장 높았으며, 발효시간 증가와 함께 증가하는 경향을 보였다. 발효 누에분말의 환원력은 B. subtilis 균주 발효에 의해서는 6일째에 A. kawachii 균주 발효에 의해서는 12일째 가장 높은 것으로 나타나 균주간에 차이가 있었다. 혈전 분해 활성은 B. subtilis 및 A. kawachii 두 균주 발효 모두 6일째에 가장 높았으며, 이러한 효과는 B. subtilis 발효 누에분말보다는 A. kawachii 발효 누에분말에서 더 높았다. SDS-PAGE상의 단백질 패턴 분석에서는 66-97 kDa 크기의 단백질 밴드가 B. subtilis 발효에 의해서는 발효 초기인 3일째 대부분 분해가 일어났으나, A. kawachii 발효 누에분말에서는 발효 3일째부터 차츰 분해되기 시작하여 12일째 분해가 대부분 이루어졌다. 따라서 미생물을 이용한 발효 누에에서 특정의 생리활성 작용을 기대하기 위해서는 사용하는 균종에 따라 발효 기간이 달라져야 한다는 것을 시사하였다.

• Biomolecules & Therapeutics
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• 제22권6호
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• pp.525-531
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• 2014

In the present study, we investigated whether apigenin significantly affects tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced production and gene expression of MUC5AC mucin in airway epithelial cells. Confluent NCI-H292 cells were pretreated with apigenin for 30 min and then stimulated with TNF-${\alpha}$ for 24 h or the indicated periods. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Apigenin significantly inhibited MUC5AC mucin production and down-regulated MUC5AC gene expression induced by TNF-${\alpha}$ in NCI-H292 cells. To elucidate the action mechanism of apigenin, effect of apigenin on TNF-${\alpha}$-induced nuclear factor kappa B (NF-${\kappa}B$) signaling pathway was also investigated by western blot analysis. Apigenin inhibited NF-${\kappa}B$ activation induced by TNF-${\alpha}$. Inhibition of inhibitory kappa B kinase (IKK) by apigenin led to the suppression of inhibitory kappa B alpha ($I{\kappa}B{\alpha}$) phosphorylation and degradation, p65 nuclear translocation. This, in turn, led to the down-regulation of MUC5AC protein production in NCI-H292 cells. Apigenin also has an influence on upstream signaling of IKK because it inhibited the expression of adaptor protein, receptor interacting protein 1 (RIP1). These results suggest that apigenin can regulate the production and gene expression of mucin through regulating NF-${\kappa}B$ signaling pathway in airway epithelial cells.

• Natural Product Sciences
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• 제12권4호
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• pp.205-209
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• 2006

The structures of flavonoids, 2(S)-5,7-dihydroxy-8,2'-dimethoxyflavanone (1), wogonin (2), 2(S)-5,7, 2'-trihydroxy-8-methoxyflavanone (3), and 2(S)-5,2',5'-trihydroxy-7,8-dimethoxyflavanone (4), isolated from Scutellaria indica were revised on the basis of 2D NMR spectroscopy, including to gCOSY, gHSQC, and gHMBC. Compounds 1-4 were tested in vitro protein tyrosine phosphatase 1B (PTP1B) inhibitory activity. Compounds 2 and 4 exhibited weak PTP1B inhibitory activity with $IC_{50}$ values of 208 and $337{\mu}M$, respectively.

• Toxicological Research
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• 제13권4호
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• pp.417-421
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• 1997

Synaptobrevin is a kind of vesicle associated membrane proteins (VAMPs) which plays a secretary role in the neuronal synapse and was recently known as the biochemical target of botulinum neurotoxin type B. The structural gene of the synaptobrevin was cloned from mouse brain using RT-PCR technique and was seqrtenced. The deduced amino acid sequence showed that the synaptobrevin protein from mouse brain is exactly the same with that of the rat brain in the amino acid level. The synaptobrevin gene was subcloned into pET3a vector and expressed in E. coli. The molecular weight of the recombinant protein was 19 kDa as expected. Moreover, when the recombinant synaptobrevin protein was incubated with the native neurotoxin of Clostridium botulinum type B, it was cleaved by the toxin in a time dependent manner. This implies that the recombinant synaptobrevin protein and the native toxin are reacted in the same way as the native synaptobrevin did in the neuronal cells.

• Archives of Pharmacal Research
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• 제24권2호
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• pp.136-143
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• 2001

Fumonisins are specific inhibitors of ceramide synthase in sphingolipid metabolism. An alteration in sphingolipid metabolism as a result of fumonisin exposure is related to cell death (Yoo et al., 1992). The objective of this study was to investigate whether elevated free sphinganine levels are related to the sensitivity of cultured cells to fumonisin exposure. Fumonisin $B_1$ elevated the intracellular free sphinganine concentraions in both LLC-$PK_1$ and Chinese hamster ovary (CHO) cells. However, CHO cells are resistant to fumonisin cytotoxicity at 50${u}m$, while LLC-$PK_1$ cells are sensitive at concentrations greater than 357M. The intracellular concentration of free sphinganine in LLC-$PK_1$ cells treated at 50${u}m$ fumonisin $B_1$ for 72 h was approximately 1450 pmol/mg protein relative to the 37 pmol observed in the control culture. Under the same conditions, the population of apoptotic cells in the 50${u}m$ fumonisin $B_1$-treated culture was approximately 37% of the total compared to 12% in the control. The caspase III-like activity after 72 h in the 50${\mu}$M fumonisin $B_1$-exposed culture Increased to approximately 50 $pmol/mg$ protein/hr compared to 6 $pmol/mg$ protein/hr in the control. L-cycloserine, a serine palmitoyltransferase inhibitory reduced the fumonisin $B_1$-stimulated caspase III-like activity down to the control level. Under the same culture conditions, the intracellular concentration of free sphinganine after-cycloserine plus fumonisin $B_1$ treatment was 140 pmol/mg protein compared to 1450 $pmol/mg$ protein in fumonisin $B_1$ alone. The intracellular concentration of free sphinganine in CHO cells treated with 50${u}m$ fumonisin $B_1$ for 72 h was al)proximately 460 pmol/mg protein, indicating that the mass amount of elevated free sphinganine in the CHO cells was about 32% of that in LLC-$PK_1$ cells. Adding exogenous sphinganine to the CHO cells along with 50${u}m$ fumonisin $B_1$ treatment for 72 h caused both necrosis and apoptosis. In conclusion, the elevated endogenous sphinganine acts as a contributing factor to the fumonisin-induced cell death.