• Clinical and Experimental Pediatrics
• /
• 제48권6호
• /
• pp.580-587
• /
• 2005

New evidence is emerging that airway smooth muscle(ASM) may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, polypeptide growth factors, extracellular matrix proteins, cell adhesion receptors and co-stimulatory molecules. ASM can promote the formation of the interstitial extracellular matrix, and potentially contribute to the alterations within the extracellular matrix in asthma. In addition, extracellular matrix components can alter the proliferative, survival, and cytoskeletal synthetic function of ASM cells through integrin-directed signaling. Increased ASM mass is one of the most important features of the airway wall remodeling process in asthma. Three different mechanisms may contribute to the increased ASM mass : cell proliferation, increased migration and decreased rate of apoptosis. The major signaling pathways of cell proliferation activated by ASM mitogens are those dependent on extracellular signal-regulated kinase and phosphoinositide 3'-kinase. The key signaling mechanisms of cell migration have been identified as the p38 mitogen-activated protein kinase and the p21-activated kinase 1 pathways. ASM cells contain ${\beta}2$-adrenergic receptors and glucocorticoid receptors. They may represent a key target for ${\beta}2$-adrenergic receptor agonist/corticosteroid interactions which have antiproliferative activity against a broad spectrum of mitogens.

• KSBB Journal
• /
• 제16권2호
• /
• pp.158-162
• /
• 2001

A halotolerant and extracellular protease-producing yeast was isolated from traditional Meju and identified as a strain of Hansenular polymorpha by investigating its microbiological characteristics. The optimum pH, temperature and NaCl concentration reauired for the growth of Hansenular polymorpha S-9 were found to be pH 6.0, 30$^{\circ}C$ and 0.5 M, respectively. Extracellular protease was produced maximally at 10 U ml(sup)-1 when Hansenular polymorpha S-9 was grown on the medium containing 1.0% beef extract and 0.1 M NaCl for 12 hr at 30$^{\circ}C$. About 13% of the angiotensin-converting enzyme (ACE) inhibitory activity was shown in the hydrolysates which were obtained from the digestion of soybean protein (6 mg) for 6 hr at 30$^{\circ}C$ by the crude enzyme (1 U).

• 한국어병학회지
• /
• 제18권3호
• /
• pp.215-225
• /
• 2005

본 연구는 강독성 균주인 E. tarda PoKF-000623가 생산하는 세포외생성물 (ECPs)을 넙치에 투여되었을 때 나타나는 생리학적 및 면역학적 영향을 조사하고자 하였다. E. tarda PoKF-000623 균주가 생성하는 ECPs의 넙치 (평균체중, 5.6g)에 대한 96hr-$LD_{50}$ 은 약 0.714${\mu}g$ g TEX>$fish^{-1}$ 이었으며, ECPs의 주사에 의한 폐사는 고농도 (40${\mu}g$ TEX>$fish^{-1}$) 뿐만 아니라 저농도 (4${\mu}g$ TEX>$fish^{-1}$)에서도 지속적으로 발생하였다. 평균 체중 59.1 g 크기의 넙치에 각각 0, 4 and 40${\mu}g$ ECPs TEX>$fish^{-1}$ 의 농도로 근육 주사한 후, 혈청 중 glucose 농도와 total protein 농도 및 신장 식세포의 활성을 조사하였다. 그 결과 고농도의 ECPs 주사 시험구의 넙치에서 혈중 glucose 농도와 total protein 농도가 대조구에 비하여 유의적으로 높게 나타났다. 그러나 세포성 면역 기구인 신장 식세포의 CL 반응과 세균 살해능은 대조구에 비하여 유의적으로 저하되었다. 따라서 넙치에 고농도의 E. tarda ECPs를 투여하면 생체 지표의 일부에 영향을 미치는 것으로 확인되었고 면역 활성, 특히 신장 식세포 활성의 억제가 edwardsielosis의 발병에 영향을 미치는 것으로 사료된다.

• Archives of Pharmacal Research
• /
• 제26권10호
• /
• pp.846-854
• /
• 2003

The oligomerization of G-proteincoupled receptors (GPCRs) has been shown to occur by various mechanisms, such as via disulfide covalent linkages, non covalent (ionic, hydrophobic) interactions of the N-terminal, and/or transmembrane and/or intracellular domains. Interactions between GPCRs could involve an association between identical proteins (homomers) or non-identical proteins (heteromers), or between two monomers (to form dimers) or multiple monomers (to form oligomers). It is believed that muscarinic receptors may also be arranged into dimeric or oigomeric complexes, but no systematic experimental evidence exists concerning the direct physical interaction between receptor proteins as its mechanism. We undertook this study to determine whether muscarinic receptors form homomers or a heteromers by direct protein-protein interaction within the same or within different subtypes using a yeast two-hybrid system. Intracellular loops (i1, i2 and i3) and the C-terminal cytoplasmic tails (C) of human muscarinic (Hm) receptor subtypes, Hm1, Hm2 and Hm3, were cloned into the vectors (pB42AD and pLexA) of a two-hybrid system and examined for heteromeric or homodimeric interactions between the cytoplasmic domains. No physical interaction was observed between the intracellular domains of any of the Hm/Hm receptor sets tested. The results of our study suggest that the Hm1, Hm2 and Hm3 receptors do not form dimers or oligomers by interacting directly through either the hydrophilic intracellular domains or the C-terminal tail domains. To further investigate extracellular domain interactions, the N-terminus (N) and extracellular loops (o1 and o2) were also cloned into the two-hybrid vectors. Interactions of Hm2N with Hm2N, Hm2o1, Hm2o2, Hm3N, Hm3o1 or Hm3o2 were examined. The N-terminal domain of Hm2 was found to have no direct interaction with any extracellular domain. From our results, we excluded the possibility of a direct interaction between the muscarinic receptor subtypes (Hm1, Hm2 and Hm3) as a mechanism for homo- or hetero-meric dimerization/oligomerization. On the other hand, it remains a possibility that interaction may occur indirectly or require proper conformation or subunit formation or hydrophobic region involvement.

• 유기물자원화
• /
• 제27권4호
• /
• pp.43-49
• /
• 2019

본 연구는 알칼리·초음파 전처리를 통한 하수슬러지의 가용화 정도를 확인하기 위해 슬러지 가용화율과 VSS 감량화율을 측정하였다. 또한 슬러지 가용화와 EPS간의 관계를 확인하기 위해 LB-EPS(Loosely-Bound EPS), TB-EPS(Tightly-Bound EPS)를 측정하였다. 실험 결과, TS 1.0%, pH 12 조건에서 슬러지 가용화율은 27.7% 증가하였고, LB-EPS as Carbohydrate와 Protein은 각각 14.6, 13.3 mg/L/g TS가 증가하여 가용화에 따른 유기물의 변화와 유사한 경향을 나타내었다. 또한 VSS는 26.7% 감량되었고, TB-EPS as Carbohydrate와 Protein은 각각 15.7, 21.9 mg/L/g TS 용출되어 가용화에 따른 고형물의 변화 역시 유사한 경향을 나타내었다.

• Molecules and Cells
• /
• 제38권2호
• /
• pp.145-150
• /
• 2015

Continuous intra- and extracellular stresses induce disorder of $Ca^{2+}$ homeostasis and accumulation of unfolded protein in the endoplasmic reticulum (ER), which results in ER stress. Severe long-term ER stress triggers apoptosis signaling pathways, resulting in cell death. Neural epidermal growth factor-like like protein 2 (NELL2) has been reported to be important in protection of cells from cell death-inducing environments. In this study, we investigated the cytoprotective effect of NELL2 in the context of ER stress induced by thapsigargin, a strong ER stress inducer, in Cos7 cells. Overexpression of NELL2 prevented ER stress-mediated apoptosis by decreasing expression of ER stress-induced C/EBP homologous protein (CHOP) and increasing ER chaperones. In this context, expression of anti-apoptotic Bcl-xL was increased by NELL2, whereas NELL2 decreased expression of pro-apoptotic proteins, such as cleaved caspases 3 and 7. This anti-apoptotic effect of NELL2 is likely mediated by extracellular signal-regulated kinase (ERK) signaling, because its inhibitor, U0126, inhibited effects of NELL2 on the expression of anti- and pro-apoptotic proteins and on the protection from ER stress-induced cell death.

• Journal of Microbiology and Biotechnology
• /
• 제17권6호
• /
• pp.897-904
• /
• 2007

The aim of this paper is to discuss the methodology of our investigation of the dynamics of protein degradation and the total in situ protealytic activity in meso/eutrophic, eutrophic, and hypereutrophic freshwater environments. Analysis of the kinetics and rates of enzymatic release of amino acids in water samples preserved with sodium azide allows determination of the concentrations of labile proteins $(C_{LAB})$, and their half-life time $(T_{1/2})$. Moreover, it gives more realistic information on resultant activity in situ $(V_{T1/2})$ of ecto- and extracellular proteases that are responsible for the biological degradation of these compounds. Although the results provided by the proposed method are general y well correlated with those obtained by classical procedures, they better characterize the dynamics of protein degradation processes, especially in eutrophic or hypereutrophic lakes. In these environments, processes of protein decomposition occur mainly on the particles and depend primarily on a metabolic activity of seston-attached bacteria. The method was tested in three lakes. The different degree of eutrophication of these lakes was clearly demonstrated by the measured real proteolytic pattern and confirmed by conventional trophic state determinants.

• 한국미생물·생명공학회지
• /
• 제47권4호
• /
• pp.546-554
• /
• 2019

This study, for the first time, reports the functional expression of lipase B derived from the yeast Candida antarctica (CALB) in Corynebacterium strain using the Escherichia coli plasmid PK18. The CALB gene fragment encoding a 317-amino-acid protein was successfully obtained from the total RNA of C. antarctica. CALB was readily produced in the Corynebacterium strain without the use of induction methods described in previous studies. This demonstrated the extracellular production of CALB in the Corynebacterium strain. CALB produced in the Corynebacterium MB001 strain transformed with pEC-CALB recombinant plasmid exhibited maximum extracellular enzymatic activity and high substrate affinity. The optimal pH and temperature for the hydrolysis of 4-nitrophenyl laurate by CALB were 9.0 and 40℃, respectively. The enzyme was stable at pH 10.7 in the glycine-KOH buffer and functioned as an alkaline lipase. The CALB activity was inhibited in the presence of high concentration of Mg²⁺, which indicated that CALB is not a metalloenzyme. These properties are key for the industrial application of the enzyme.

• The Korean Journal of Physiology and Pharmacology
• /
• 제22권2호
• /
• pp.193-201
• /
• 2018

Connective tissue growth factor (CTGF) is a novel fibrotic mediator, which is considered to mediate fibrosis through extracellular matrix (ECM) synthesis in diabetic cardiovascular complications. Statins have significant immunomodulatory effects and reduce vascular injury. We therefore examined whether fluvastatin has anti-fibrotic effects in vascular smooth muscle cells (VSMCs) and elucidated its putative transduction signals. We show that advanced glycation end products (AGEs) stimulated CTGF mRNA and protein expression in a time-dependent manner. AGE-induced CTGF expression was mediated via ERK1/2, JNK, and Egr-1 pathways, but not p38; consequently, cell proliferation and migration and ECM accumulation were regulated by CTGF signaling pathway. AGE-stimulated VSMC proliferation, migration, and ECM accumulation were blocked by fluvastatin. However, the inhibitory effect of fluvastatin was restored by administration of CTGF recombinant protein. AGE-induced VSMC proliferation was dependent on cell cycle arrest, thereby increasing G1/G0 phase. Fluvastatin repressed cell cycle regulatory genes cyclin D1 and Cdk4 and augmented cyclin-dependent kinase inhibitors p27 and p21 in AGE-induced VSMCs. Taken together, fluvastatin suppressed AGE-induced VSMC proliferation, migration, and ECM accumulation by targeting CTGF signaling mechanism. These findings might be evidence for CTGF as a potential therapeutic target in diabetic vasculature complication.

• 한국균학회지
• /
• 제26권4호통권87호
• /
• pp.496-501
• /
• 1998

송이균사(Tricholoma matsutake DGUM 26001, DGUM 26101, DGUM 26210, FRI 91024)를 사용하여 균사의 효소활성을 측정하였다. $24^{\circ}C$에서 30일간 배양후 그 여액을 조효소 용액으로 사용하였을 때, ${\alpha}-amylase$ 효소의 평균 비활성은 6142.3 unit/mg protein이었다. 배양여액 중의 xylanase의 세포의 효소활성은 상대적으로 높았으나, ${\beta}-glucosidase$, ligninase, CMCase, chitinase, pretense및 lipase의 효소활성은 낮거나 거의 없었다.