• Title/Summary/Keyword: extracellular protein

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Proliferative and Synthetic Responses of Airway Smooth Muscle in Asthma (천식에서 기도평활근의 증식과 합성 반응에 대한 최신지견)

  • Shim, Jung Yeon
    • Clinical and Experimental Pediatrics
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    • v.48 no.6
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    • pp.580-587
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    • 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.

Isolation of a Halotolerant Yeast and the Production of Extracellular Protease (내염성 효모의 분리 및 세포외 Protease의 생산)

  • 정승찬;현광욱;김재호;이종수
    • KSBB Journal
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    • v.16 no.2
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    • pp.158-162
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    • 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).

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Effect of extracellular products (ECPs) of Edwardsiella tarda on olive flounder, Paralichthys olivaceus (Edwardsiella tarda extracellular products (ECPs)의 인위투여에 따른 넙치, Paralichthys olivaceus의 생체반응)

  • Lee, Deok-Chan;Kim, Yi-Cheong;Kim, Jin-Woo;Park, Soo-Il
    • Journal of fish pathology
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    • v.18 no.3
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    • pp.215-225
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    • 2005
  • The effect of extracellular products (ECPs) prepared from highly virulent Edwardsiella tarda PoKF-000623 on the physiological and the immunological function in the olive flounder, Paralichthys olivaceus was evaluated. The virulence of ECPs on the fish, 5.6 g of average body weight was detected $0.714{\mu}{g}$ g $fish^{-1}$ as $96hr-LD_{50}$ by intraperitonial injection. After intramuscularly injected with 0, 4 and $40{\mu}{g}$ ECPs TEX>$fish^{-1}$ to fish average sized 59.1 g of body weight, the fish were measured the glucose and total protein concentration in sera, and immune activity of kidney macrophage. The fish injected with $40{\mu}{g}$ protein $fish^{-1}$ showed a significant increase in the glucose and total protein concentration in sera. But, the fish were changed in cellular immune response as lowed chemiluminescence response and bactericidal activity of head kidney macrophage. These results were suggested that ECPs of E. tarda could be effect on the physiological and the immunological factors, especially in the function of kidney macrophage concerning to edwardsiellosis infection.

Homo- or Hetero-Dimerization of Muscarinic Receptor Subtypes is Not Mediated by Direct Protein-Protein Interaction Through Intracellular and Extracellular Regions

  • Kang, Yun-Kyung;Yoon, Tae-Sook;Lee, Kyung-Lim;Kim, Hwa-Jung
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.846-854
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    • 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.

The relation between sewage sludge solubilization and extracellular polymeric substances (하수슬러지 가용화와 체외고분자물질(EPS)간의 관계)

  • Jeon, Byeong-Cheol;Nam, Se-Yong
    • Journal of the Korea Organic Resources Recycling Association
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    • v.27 no.4
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    • pp.43-49
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    • 2019
  • In order to investigate the relation between sewage sludge solubilization and extracellular polymeric substances(EPS) during alkaline-ultrasonic pretreatment, SCOD/TCOD ratio, solubilization rate, VSS/TS ratio, VSS reduction rate, LB-EPS(Loosely-Bound EPS) and TB-EPS(Tightly-Bound EPS) were measured. At the condition of TS 1.0% and pH 12, solubilization rate increased by 27.7%, LB-EPS as Carbohydrate and Protein increased by 14.6 and 13.3 mg/L/g TS, respectively. Withal, VSS decreased by 26.7% and TB-EPS as Carbohydrate and Protein were extracted by 15.7 and 21.9 mg/L/g TS, respectively. Consequently, the concentrations of organic matter and LB-EPS increased and the trends appeared similarly. In addition, the concentrations trend of decreasing solid matter and extracted TB-EPS also appeared similarly.

NELL2 Function in the Protection of Cells against Endoplasmic Reticulum Stress

  • Kim, Dong Yeol;Kim, Han Rae;Kim, Kwang Kon;Park, Jeong Woo;Lee, Byung Ju
    • Molecules and Cells
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    • v.38 no.2
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    • pp.145-150
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    • 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.

The Dynamics of Protein Decomposition in Lakes of Different Trophic Status - Reflections on the Assessment of the Real Proteolytic Activity In Situ

  • Siuda, Waldemar;Kiersztyn, Bartosz;Chrost, Ryszard J.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.897-904
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    • 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.

Molecular Cloning and Expression of Candida antarctica lipase B in Corynebacterium genus

  • Gonzalez, Tamara;M'Barek, Hasna Nait;Gomaa, Ahmed E.;Hajjaj, Hassan;Zhen, Chen;Dehua, Liu
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.546-554
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    • 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.

Fluvastatin inhibits advanced glycation end products-induced proliferation, migration, and extracellular matrix accumulation in vascular smooth muscle cells by targeting connective tissue growth factor

  • Hwang, Ae-Rang;Nam, Ju-Ock;Kang, Young Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.2
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    • pp.193-201
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    • 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.

The Extracellular Enzyme Activities in Culture Broth of Tricholoma matsutake (송이균사(Tricholoma matsutake) 배양액의 세포외 효소 활성)

  • Lee, Chang-Yun;Hong, Oun-Pyo;Jung, Myung-Jun;Han, Yeong-Hwan
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.496-501
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    • 1998
  • The mycelia of Tricholoma matsutake DGUM 26001, 26101, 26210 and FRI 91024 were used to determine the extracellular enzyme activity in mycelia. When the filtrate of culture broth after 30-day cultivation at $24^{\circ}C$ was used as a crude solution of extracellular enzyme, the average specific activity of ${\alpha}-amylase$ was 6142.3 unit/mg protein. The specific activity of xylanase was comparatively high. However, little or no enzyme activities were found for ${\beta}-glucosidase$, ligninase, CMCase, chitinase, protease, and lipase.

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