• Title/Summary/Keyword: Functional compartment

Search Result 31, Processing Time 0.025 seconds

Endoplasmic Reticulum Signaling for Recombinant-protein Production (재조합 단백질 생산을 위한 소포체 신호전달)

  • Goo, Tae-Won;Yun, Eun-Young;Kang, Seok-Woo;Kwon, Ki-Sang;Kwon, O-Yu
    • Journal of Life Science
    • /
    • v.17 no.6 s.86
    • /
    • pp.847-858
    • /
    • 2007
  • The endoplasmic reticulum (ER) is an important intracellular organelle for folding and maturation of newly synthesized transmembrane and secretory proteins. The ER provides stringent quality control systems to ensure that only correctly folded proteins exit the ER and unfolded or misfolded proteins are retained and ultimately degraded. The ER has evolved stress response both signaling pathways the unfolded protein response (UPR) to cope with the accmulation of unfolded or misfolded proteins and ER overload response (EOR). Accumulating evidence suggests that, in addition to responsibility for protein processing, ER is also an important signaling compartment and a sensor of cellular stress. In this respect, production of bio-functional recombinant-proteins requires efficient functioning of the ER secretory pathway in host cells. This review briefly summarizes our understanding of the ER signaling developed in the recent years to help of the secretion capacities of recombinant cells.

Effect of a Refrigerator with LED on Functional Composition Changes and Freshness Prolongation of Cabbage (냉장고의 LED부착이 양배추의 선도 연장에 미치는 영향)

  • Park, Shin-Young;Chang, Min-Sun;Choi, Jung-Hee;Kim, Byeong-Sam;Lee, Hye-Ran;Ham, Kyung-Hee
    • Food Science and Preservation
    • /
    • v.14 no.2
    • /
    • pp.113-118
    • /
    • 2007
  • A LED (light emitting diode) was attached inside the cold $(4^{\circ}C)$ compartment of a home refrigerator to investigate a possible effect on freshness prolongation of cabbage over 10 d of storage. The LED increased chlorophyll synthesis, inhibited vitamin C degradation and increased the content of phenolics, compared to values measured in cabbages stored in an identical refrigerator without the LED. In the refrigerator with the LED, the color of cabbage leaves remained green for a period fourfold longer than that seen in a conventional refrigerator. Vitamin C content was twice that of a conventionally stored cabbage, and phenolic consent was 14% higher in the LED-illuminated vegetables.

Molecular and Biochemical Properties of a Cysteine Protease of Acanthamoeba castellanii

  • Hong, Yeonchul;Kang, Jung-Mi;Joo, So-Young;Song, Su-Min;Le, Huong Giang;Thai, Thl Lam;Lee, Jinyoung;Goo, Youn-Kyoung;Chung, Dong-Il;Sohn, Woon-Mok;Na, Byoung-Kuk
    • Parasites, Hosts and Diseases
    • /
    • v.56 no.5
    • /
    • pp.409-418
    • /
    • 2018
  • Acanthamoeba spp. are free-living protozoa that are opportunistic pathogens for humans. Cysteine proteases of Acanthamoeba have been partially characterized, but their biochemical and functional properties are not clearly understood yet. In this study, we isolated a gene encoding cysteine protease of A. castellanii (AcCP) and its biochemical and functional properties were analyzed. Sequence analysis of AcCP suggests that this enzyme is a typical cathepsin L family cysteine protease, which shares similar structural characteristics with other cathepsin L-like enzymes. The recombinant AcCP showed enzymatic activity in acidic conditions with an optimum at pH 4.0. The recombinant enzyme effectively hydrolyzed human proteins including hemoglobin, albumin, immunoglobuins A and G, and fibronectin at acidic pH. AcCP mainly localized in lysosomal compartment and its expression was observed in both trophozoites and cysts. AcCP was also identified in cultured medium of A. castellanii. Considering to lysosomal localization, secretion or release by trophozoites and continuous expression in trophozoites and cysts, the enzyme could be a multifunctional enzyme that plays important biological functions for nutrition, development and pathogenicity of A. castellanii. These results also imply that AcCP can be a promising target for development of chemotherapeutic drug for Acanthamoeba infections.

Time-dependent proteomic and genomic alterations in Toll-like receptor-4-activated human chondrocytes: increased expression of lamin A/C and annexins

  • Ha, Seung Hee;Kim, Hyoung Kyu;Nguyen, Thi Tuyet Anh;Kim, Nari;Ko, Kyung Soo;Rhee, Byoung Doo;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.21 no.5
    • /
    • pp.531-546
    • /
    • 2017
  • Activation of Toll-like receptor-4 (TLR-4) in articular chondrocytes increases the catabolic compartment and leads to matrix degradation during the development of osteoarthritis. In this study, we determined the proteomic and genomic alterations in human chondrocytes during lipopolysaccharide (LPS)-induced inflammation to elucidate the underlying mechanisms and consequences of TLR-4 activation. Human chondrocytes were cultured with LPS for 12, 24, and 36 h to induce TLR-4 activation. The TLR-4-induced inflammatory response was confirmed by real-time PCR analysis of increased interleukin-1 beta ($IL-1{\beta}$), interleukin-6 (IL-6), and tumor necrosis factor alpha ($TNF-{\alpha}$) expression levels. In TLR-4-activated chondrocytes, proteomic changes were determined by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization-mass spectroscopy analysis, and genomic changes were determined by microarray and gene ontology analyses. Proteomics analysis identified 26 proteins with significantly altered expression levels; these proteins were related to the cytoskeleton and oxidative stress responses. Gene ontology analysis indicated that LPS treatment altered specific functional pathways including 'chemotaxis', 'hematopoietic organ development', 'positive regulation of cell proliferation', and 'regulation of cytokine biosynthetic process'. Nine of the 26 identified proteins displayed the same increased expression patterns in both proteomics and genomics analyses. Western blot analysis confirmed the LPS-induced increases in expression levels of lamin A/C and annexins 4/5/6. In conclusion, this study identified the time-dependent genomic, proteomic, and functional pathway alterations that occur in chondrocytes during LPS-induced TLR-4 activation. These results provide valuable new insights into the underlying mechanisms that control the development and progression of osteoarthritis.

Effect of the Contents Ratio of Panaxadiol Ginsenosides Extracted from Various Compartment of Ginseng on the Transcription of Cu/Zn Superoxide Dismutase Gene (홍삼의 각 부위에서 추출된 Panaxadiol분획의 함량비에 따른 유해산소제거효소(Cu/Zn Superoxide Dismutase) 유도효과)

  • Chang Mun Seog;Choi Kang Ju;Rho Hyune Mo
    • Journal of Ginseng Research
    • /
    • v.23 no.1 s.53
    • /
    • pp.44-49
    • /
    • 1999
  • Cu/Zn superoxide dismutase (SOD1) is a protective enzyme responsible for the dismutat ion of superoxide radicals within the cell by converting superoxide radicals to oxygen and hydrogen peroxide, which is in turn changed to oxygen and water by catalase. Previously, we reported that the panaxadiol (PD) and its ginsenoside $Rb_2$ induced the expression of SOD1 gene through AP2 binding site and its induction. Here, we examined the effect of subfractions of panaxadiol ginsenosides, which were extracted from different parts of ginseng root that possess various ratios of panaxadiol to panaxatriol, on the induction of SOD1 gene expression. To explore this possibility, the upstream regulatory region of SOD1 was linked to the chloramphenicol acetyl transferase (CAT) structural gene and introduced into human hepatoma HepG2 cells. We observed that the transcriptional activation of SOD1 was proportional to the contents ratio of panaxadiol ginsensides. Consistent with this results, the total extract portion prepared from the finely-hairy root, which contains the higher ratio of panaxadiol to panaxatriol about 2.6, increased the SODl transcription about 3 fold. This results suggest that the panaxadiol fraction could induce the SOD1 and total extract of the ginseng finely-hairy root would be a useful material as a functional food for the SOD1 inducer.

  • PDF

Formulation and Skin Penetration Characteristics of Aceclofenac Plaster for Transdermal Delivery (아세클로페낙의 경피 제제설계 및 피부투과 특성)

  • Chung, Jong-Keun;Lee, Min-Suk;Park, Jeong-Hwa;Lee, Jang-Won;Kim, Ha-Hyung;Choi, Young-Wook;Lee, Kwang-Pyo
    • Journal of Pharmaceutical Investigation
    • /
    • v.29 no.1
    • /
    • pp.29-36
    • /
    • 1999
  • Aceclofenac is an non-steroidal antiinflammatory drug which has been used in the treatment of rheumatoidal rthritis and osteo-arthritis. In order to decrease the gastric ulcerogenic effects and contol the plasma level of aceclofenac, we have developed the transdermal delivery system of aceclofenac plaster, which were formulated employing matrix polymers of acrylates and penetration-enhancers such as $Lauroglycol^{\circledR}$, $Transcutol^{\circledR}$, oleic acid and linoleic acid. Using Franz diffusion cells mounted with a rat skin, transdermal penetration characteristics of the formulations were evaluated by the HPLC assay of aceclofenac and diclofenac, an active metabolite, in the receptor compartment of pH 7.2 phosphate buffered solution. Skin penetration was increased when the content of aceclofenac increased, showing the flux $(J,\;{\mu}g/cm^2/hr)$ of 0.37 and 2.50 for 2% and 6.75% of the content, respectively. The flux$(J,\;{\mu}g/cm^2/hr)$ from plasters made of $Durotak^{\circledR}$ 87-2074, $Durotak^{\circledR}$ 87-2510 and $Durotak^{\circledR}$ 87-2097 were 2.50, 2.77 and 4.39, respectively. $Durotak^{\circledR}$ 87-2074 showed the lowest penetration due to the carboxylic acid group in the polymer, which might form a strong hydrogen bonding with a secondary amine of aceclofenac. Although both $Durotak^{\circledR}$ 87-2510 and $Durotak^{\circledR}$ 87-2097 are amine-resistant adhesives, $Durotak^{\circledR}$ 872510 showed lower penetration than $Durotak^{\circledR}$ 87-2097 because of the hydroxyl group in $Durotak^{\circledR}$ 87-2510, which might form a weak hydrogen bonding with aceclofenac. These results reveal that the functional group in acrylic polymers would greatly affect the release of aceclofenac from the matrix, which is the rate-limiting step in the penetration of aceclofenac through rat skins. The penetration of aceclofenac from plasters using different penetration-enhancers increased in the following order: Transcutol < linoleic acid < oleic acid. And the flux from the plasters containing oleic acid as a penetrationenhancer was 2.22 times greater than that of creams, which suggest that a newly deveolped aceclofenac plaster could be used in the treatment of rheumatoidal arthritis and osteo-arthritis as an advanced transdermal delivery system.

  • PDF

Subcellular partitioning-dependent functional switching of Arabidopsis photoreceptor phytochrome B in response to brassinosteroids

  • Ryu, Jong-Sang;Choi, Hyun-Mo;Hong, Sung-Hyun;Matsushita, Tomonao;Nagatani, Akira;Nam, Hong-Gil
    • Interdisciplinary Bio Central
    • /
    • v.1 no.1
    • /
    • pp.1.1-1.5
    • /
    • 2009
  • Many organisms control their physiology and behavior in response to the local light environment, which is first perceived by photoreceptors that undergo light-dependent conformational changes. Phytochromes are one of the major photoreceptors in plants, controlling wide aspects of plant physiology by recognizing the light in red (R) and far-red (FR) spectra. Higher plants have two types of phytochromes; the photo-labile type I (phyA in Arabidopsis) and photo-stable type II (phyB-E in Arabidopsis). Phytochrome B (phyB), a member of the type II phytochromes in Arabidopsis, shows classical R and FR reversibility between the inter-convertible photoisomers, Pr and Pfr. Interestingly, the Pr and Pfr isomers show partitioning in the cytosol and nucleus, respectively. In the over 50 years since its discovery, it has been thought that the type II phytochromes only function to mediate R light. As described in the text, we have now discovered phyB has an active function in FR light. Even striking is that the R and FR light exert an opposite effect. Thus, FR light is not simply nullifying the R effect but has an opposing effect to R light. What is more interesting is that the phyB-mediated actions of FR and R light occur at different cellular compartment of the plant cell, cytosol and nucleus, respectively, which was proven through utilization of the cytosolic and nuclear-localized mutant versions of phyB. Our observations thus shoot down a major dogma in plant physiology and will be considered highly provocative in phytochrome function. We argue that it would make much more sense that plants utilize the two isoforms rather than only one form, to effectively monitor the changing environmental light information and to incorporate the information into their developmental programs.

Functional Analysis of a Grapevine UDP-Glucose Flavonoid Glucosyl Transferase (UFGT) Gene in Transgenic Tobacco Plants (담배 형질전환체를 이용한 포도 UDP-glucose flavonoid glucosyl transferase (UFGT) 유전자의 기능 분석)

  • Park, Ji-Yeon;Park, Sung-Chool;Pyee, Jae-Ho
    • Journal of Life Science
    • /
    • v.20 no.2
    • /
    • pp.292-297
    • /
    • 2010
  • Anthocyanin, a phenolic compound, is a pigment that shows blue or red color in the fruit, petal and other tissues. It is an important factor in grape berry skin pigment and accumulates only in the skin. This skin-specific accumulation of anthocyanin has been reported to be regulated by the ufgt gene which encodes UDP-glucose: flavonoid 3-O-glucosyltransferase that participates in the biosynthesis of anthocyanin. The ufgt gene is expressed only in berry skin, while the other genes involved in the biosynthetic pathway are expressed in both skin and flesh tissues. In order to determine whether anthocyanin accumulation is primarily regulated by compartment of UFGT, a ufgt cDNA clone was isolated from grape berry, its open reading frame was ligated in pBI121 vector in either a sense or an antisense orientation under the control of the CaMV35S promoter and the recombinant constructs were incorporated into tobacco plants. Several transgenic lines were selected and characterized to determine the level of expression of the grapevine ufgt transcript and endogenous homologs of tobacco. Compared to the wild-type, the amount of anthocyanins in sense transgenic plants increased by 44%, while the amount of anthocyanins in antisense transgenic plants decreased by 88%. In addition, the color of flowers became intense in the sense transgenic plants. These results suggest that over-expression or repression of the ufgt gene affected the accumulation of anthocyanin in flowers of tobacco.

MR Study of Wate Exchange and Cell Membrane Permeability in Rat Liver Cells Using a Tissue-Specific MR Contrast Agent (조직 특성 MR 조영제를 이용한 쥐의 간세포막의 물분자 교환 및 투과율의 MR 측정기법)

  • Yongmin Chang
    • Investigative Magnetic Resonance Imaging
    • /
    • v.2 no.1
    • /
    • pp.73-82
    • /
    • 1998
  • Purpose : A precise NMR technique for measuring the rate of water exchange and cell membrane permeability across the hepatocyte membrane using liver-specific MR contrast agent is described. Materials and Methods : The rat hepatocytes isolated by perfusion of the livers were used for the NMR measurements. All experiments were performed on an IBM field cycling relaxometer operating from 0.02MHz to 60 MHz proton Larmor frequency. spin-echo pulse sequence was empolyed to measure spin-lattice relaxation time, T1. The continuous distribution analysis of water proton T1 data from rat hepatocytes containing low concentrations of the liver specific contrast agent, Gd-EOB-DTPA, modeled by a general two compartment exchange model. Results : The mean residence time of water molecule inside the hepatocyte was approximately 250 msec. The lower limit for the permeability of the hepatocyte membrane was $(1.3{\pm}0.1){\;}{\times}{\;}10^{-3}cm/sec$. The CONTIN analysis, which seeks the natural distribution of relaxation times, reveals direct evidence of the effect of diffusive exchange. the diffusive water exchange is not small in the intracellular space in the case of hepatocytes. Conclusions : Gd-EOB-DTPA, when combined with continuous distribution analysis, provides a robust method to study water exchange and membrane permeability in hepatocytes. Water exchange in hepatocyte is much slower thatn that in red blood cells. Therefore, tissue-specific contrast agent may be used as a functional agent to give physiological information such as cell membrane permeability.

  • PDF

Treatment of Pyogenic Arthritis of the Knee Using Drainage Tube Insertion in the Posterior Compartment through the Posterior Transeptal Portal after an Arthroscopic Synovectomy (관절경적 활액막 절제술 후 후격막 삽입구를 통한 배액관의 후방 거치를 이용한 무릎의 화농성 관절염 치료)

  • Kim, Tae Ho;Yu, Chang Eon;Shin, Chung Shik
    • Journal of the Korean Orthopaedic Association
    • /
    • v.54 no.3
    • /
    • pp.269-275
    • /
    • 2019
  • Purpose: This study analyzed the treatment effects of drain insertion in the posteromedial portal after an arthroscopic synovectomy and posterior septum resection for pyogenic arthritis. Materials and Methods: From June 2005 to December 2016, 56 cases (55 patients, 1 case of bilateral knee) were diagnosed with pyogenic arthritis and arthroscopic treatment was performed. Fourteen patients (25.0%) were identified with causative organisms. The average follow-up period was 12.3 months, and the mean age was 67.8 years old. Twenty-four cases were males and 31 cases were females. Kellgrene-Lawrence grade (K-L grade) I was 6 patients, II was 20 patients, III was 11 patients, and IV was 18 patients. Surgery was performed through six portal. The posterior septum was removed and a drainage tube was inserted from the posterior medial side to the posterior side of the joint. The Lysholm score was used after surgery for a functional evaluation. The K-L grade at the time of admission and at the last follow-up were compared for a radiological evaluation. Results: The mean normalization period of the C-reactive protein was 59.8 days (6-164 days). Intravenous antibiotics were used for an average of 37.1 days. The results of the functional evaluation revealed a mean Lysholm score of 64.5 (30-98) in a total of 56 cases. Two reoperations was performed, but the treatment was completed with same method in all patients. The K-L grade was similar at the time of surgery and at the last follow-up (p>0.05). Conclusion: The method of drain insertion in the posteromedial portal after arthroscopic synovectomy and posterior septum resection for pyogenic arthritis is an effective technique that can obtain satisfactory results by inducing proper exudate discharge.