• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.116-122
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• 2002

The partial 165 rDNA sequences of 6 lactic acid bacterial strains isolated from Kimchi were determined. Two strains were Leuconostoc mesenteroides and the rest were incorrectly classified and turned out to be Lactobacillus. As the case of dairy lactic acid bacteria, the strains isolated from Kimchi also had cell-envelope proteinase (CEP) activity. As the result of partial CEP gene amplification with CEP-specific primers, the expected 1.2-kb amplificate was obtained not from Leu. mesenteroides but from Lactobacillus strains. The deduced amino acid sequence of PCR product amplified from the genomic DNA of Lactobacillus pentosus KFR1821 showed 95％ and 92％ homology with those of PrtPs from Lactococcus lactis subsp. cremoris and Lactobacillus paracasei subsp. paracasei, respectively. The PCR amplificate was used as a probe and the result of Southern hybridization illuminated the location of CEP gene in chromosomal DNA of Lb. pentosus KFR1821.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.470-478
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• 2012

Recent genome comparisons of E. coli B and K-12 strains have indicated that the makeup of the cell envelopes in these two strains is quite different. Therefore, we analyzed and compared the envelope proteomes of E. coli BL21(DE3) and MG1655. A total of 165 protein spots, including 62 nonredundant proteins, were unambiguously identified by two-dimensional gel electrophoresis and mass spectrometry. Of these, 43 proteins were conserved between the two strains, whereas 4 and 16 strain-specific proteins were identified only in E. coli BL21(DE3) and MG1655, respectively. Additionally, 24 proteins showed more than 2-fold differences in intensities between the B and K-12 strains. The reference envelope proteome maps showed that E. coli envelope mainly contained channel proteins and lipoproteins. Interesting proteomic observations between the two strains were as follows: (i) B produced more OmpF porin with a larger pore size than K-12, indicating an increase in the membrane permeability; (ii) B produced higher amounts of lipoproteins, which facilitates the assembly of outer membrane ${\beta}$-barrel proteins; and (iii) motility- (FliC) and chemotaxis-related proteins (CheA and CheW) were detected only in K-12, which showed that E. coli B is restricted with regard to migration under unfavorable conditions. These differences may influence the permeability and integrity of the cell envelope, showing that E. coli B may be more susceptible than K-12 to certain stress conditions. Thus, these findings suggest that E. coli K-12 and its derivatives will be more favorable strains in certain biotechnological applications, such as cell surface display or membrane engineering studies.

• The Journal of Korean Medicine
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• v.18 no.1
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• pp.430-445
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• 1997

In order to know the effects of Calculus $Bovis{\cdot}F디$ Ursi aqua-acupuncture ($C{\cdot}F$ aqua-acupuncture) to the liver damage, we carried out the treatment of $C{\cdot}F$ aqua-acupuncture and injection of Kam-Du-Tang on the liver damaged rat which is induced by aconitine, and then ultrastructural study has been to the liver tissue. The results were as follows: 1. In the hepatic cell of normal group, the nuclear envelope is round and electro-density of nucleoplasm is relatively low, and have one or two large nucleolus. Cell organelle is developed rough endoplasmic reticulum(RER) and many mitochondria, glycogen granules, smooth endoplasmic reticulum(SER) and Golgi apparatus. 2. In 24 hours control group, nuclear envelope is very irregular, RER and most of cell organell is shown that destroyed by aconitine. This is similar to the 48 hours control group. In 72 hours control group, RER is recovered somewhat but cristae of mitochondria is not seen the shape 3.In 24 hours $C{\cdot}F$ aqua-acupuncture group, nuclear envelope is very irregular and cristernae is very dilatable and many mitochondria is dilationed and the cristae is not definite like the control group. In 48 hours $C{\cdot}F$ aqua-acupuncture group, nuclear envelope is relatively round and chromatin is regular. Cristernae of RER and ribosme is somewhat imperfect but is similar the normal group, and the cristae of mitchondria is shown definitely like the normal group. In 72 hours $C{\cdot}F$ aqua-acupuncture group, nuclear envelope is round and cell organell is similar to normal group. 4. In 24 hours Kam-Du-Tang group, nuclear envlope is very irregular and cell organelles are destroyed by the effects of aconitine. In 48 hours Kam-Du-Tang group, nuclear envelope is somewhat irregular but chromatin is relatively regular. RER is seperated regularly through the cytoplasm, but cannot make the structure of lamina, and cistennae is very dilationed. many of glycogen granules is seperated regularly. In 72 hours Kam-Du-Tang group, RER makes the lamina. Mitochondria is a little but inner space is dilationed so the shape of is not definite and a lot of glycogen granules are accumulated in several places. It see the above result, hepatic cell that is damaged by aconitine is effected chiefly by the $C{\cdot}F$ acupuncture and effect of acupuncture is similar to the detoxicated effect of Kam-Du-Tang.

• Biomolecules & Therapeutics
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• v.20 no.2
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• pp.171-176
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• 2012

HaCaT cells are the immortalized human keratinocytes and have been extensively used to study the epidermal homeostasis and its pathophysiology. T helper cells play a role in various chronic dermatological conditions and they can affect skin barrier homeostasis. To evaluate whether HaCaT cells can be used as a model cell system to study abnormal skin barrier development in various dermatologic diseases, we analyzed the gene expression profile of epidermal differentiation markers of HaCaT cells in response to major T helper (Th) cell cytokines, such as $IFN{\gamma}$, IL-4, IL-17A and IL-22. The gene transcriptional profile of cornified envelope-associated proteins, such as filaggrin, loricrin, involucrin and keratin 10 (KRT10), in HaCaT cells was generally different from that in normal human keratinocytes (NHKs). This suggests that HaCaT cells have a limitation as a model system to study the pathophysiological mechanism associated with the Th cell cytokine-dependent changes in cornified envelope-associated proteins which are essential for normal skin barrier development. In contrast, the gene transcription profile change of human ${\beta}2$-defensin (HBD2) in response to $IFN{\gamma}$, IL-4 or IL-17A in HaCaT cells was consistent with the expression pattern of NHKs. $IFN{\gamma}$ also up-regulated transglutaminase 2 (TGM2) gene transcription in both HaCaT cells and NHKs. As an alternative cell culture system for NHKs, HaCaT cells can be used to study molecular mechanisms associated with abnormal HBD2 and TGM2 expression in response to $IFN{\gamma}$, IL-4 or IL-17A.

• The Journal of Korean Society of Virology
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• v.29 no.3
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• pp.183-193
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• 1999

Human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein is synthesized as a 160 KDa precursor, gp160, that is cleaved by a cellular protease to form the gp120 and gp41 subunits. Mammalian expression vectors were designed that are capable of efficient expression of various mutant envelope glycoproteins derived from a molecular clone of HIV-1. To construct these vectors, one type of mutation was made at the gp120-gp41 cleavage site by oligonucleotide-directed mutagenesis. And another mutation was made to change amino acids in the membrane spanning region of HIV-1 gp41 important for membrane anchorage. Next, these two mutations were combined to generate a vector to have double mutations in cleavage site and membrane-spanning region. These mutants were transiently expressed in mammalian cells. The effect of these mutations on envelope glycoprotein synthesis, proteolytic processing and secretion was determined. In addition, cell surface expression and ability of the glycoprotein to induce syncytium formation were examined. This study provides a mammalian expression system that is capable of efficient expression and secretion of soluble gp160.

• KIEAE Journal
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• v.4 no.3
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• pp.113-119
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• 2004

As Building Integrated Photovoltaic(BIPV) system replaces the conventional building finishing materials with PV modules, two function of electricity generation and building envelope can be expected. Therefore BIPV can be a good alternative technology for the 21 century environment-friendly buildings. The objective of this paper is to develope BIPV modules for a commercial buildings of which structure is mainly light-weight, curtain wall system. Two types of module are developed for a opaque part and a transparent part of building envelope. Current technology level and market status of Korea determines the configuration of developed BIPV modules. Architectural considerations for the integration of PV module to building envelope such as building structure, construction type, safety, regulation, maintenance etc. have been carefully reflected from the early stage of BIPV module design. Especially the survey result of current building envelope materials determines the size of unit BIPV modules and a unique cladding method for PV module installation is developed. Trial product of BIPV modules and cladding hardwares are manufactured and sample construction details for a demonstration building are proposed.

• Journal of Periodontal and Implant Science
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• v.32 no.2
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• pp.351-360
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• 2002

In the study presented here, identification, purification, and partial characterization of a hemin-regulated protein in Prevotella nigrescens were carried out. The results of this study confirm that the availability of hemin influences the expression of a selected membrane protein as well as the growth rate of P. nigrescens ATCC 33563. The 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin-binding protein from P. nigrescens. Disulfide bonds were not present in this protein, and N'-terminal amino acid sequence analysis revealed that this protein belongs to a new, so far undescribed protein. The 50 kDa protein was found to be rich in hydrophilic amino acids, with glycine comprising approximately 60% of the total amino acids. The study described here is the first to identify, purify, and biochemically characterize a putative hemin-binding protein from P. nigrescens. Work is in progress to further characterize the molecular structure of this protein.

• Journal of Periodontal and Implant Science
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• v.36 no.1
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• pp.155-165
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• 2006

The results of this study confirm that the availability of hemin influences the expression of selected membrane proteins of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is hemin regulated was identified and purified in P. gingivalis. A strong hemin-binding function was found by LDS-PAGE and TMBZ staining when P. gingivalis cells were grown under hemin-limited conditions. A 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin binding protein from P. intermedia and P. nigrescens, respectively. N-terminal amino acid sequence analysis of CNBr-digested 24 kDa hemin binding protein from P. gingivalis revealed that this protein belongs to a new, so far undescribed hemin-binding class of proteins. N-terminal amino acid sequence of a 50 kDa putative hemin binding protein from P. intermedia was identical with Enolase from Streptococcus intermedia. Work is in progress to further characterize the molecular structure of these proteins.

• Journal of Microbiology
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• v.45 no.1
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• pp.41-47
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• 2007

The Hantaan virus (HTNV) is an enveloped virus that is capable of inducing low pH-dependent cell fusion. We molecularly cloned the viral glycoprotein (GP) and nucleocapsid (NP) cDNA of HTNV and expressed them in Vero E6 cells under the control of a CMV promoter. The viral gene expression was assessed using an indirect immunofluorescence assay and immunoprecipitation. The transfected Vero E6 cells expressing GPs, but not those expressing NP, fused and formed a syncytium following exposure to a low pH. Monoclonal antibodies (MAbs) against envelope GPs inhibited cell fusion, whereas MAbs against NP did not. We also investigated the N-linked glycosylation of HTNV GPs and its role in cell fusion. The envelope GPs of HTNV are modified by N-linked glycosylation at five sites: four sites on G1 (N134, N235, N347, and N399) and one site on G2 (N928). Site-directed mutagenesis was used to construct eight GP gene mutants, including five single N-glycosylation site mutants and three double-site mutants, which were then expressed in Vero E6 cells. The oligosaccharide chain on residue N928 of G2 was found to be crucial for cell fusion after exposure to a low pH. These results suggest that G2 is likely to be the fusion protein of HTNV.

• Molecules and Cells
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• v.39 no.9
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• pp.669-679
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• 2016

N-acetyl-D-glucosamine kinase (GlcNAc kinase or NAGK) primarily catalyzes phosphoryl transfer to GlcNAc during amino sugar metabolism. Recently, it was shown NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) and upregulates axo-dendritic growth, which is an enzyme activity-independent, non-canonical structural role. The authors examined the distributions of NAGK and NAGK-dynein complexes during the cell cycle in HEK293T cells. NAGK was expressed throughout different stages of cell division and immunocytochemistry (ICC) showed NAGK was localized at nuclear envelope, spindle microtubules (MTs), and kinetochores (KTs). A proximity ligation assay (PLA) for NAGK and DYNLRB1 revealed NAGK-dynein complex on nuclear envelopes in prophase cells and on chromosomes in metaphase cells. NAGK-DYNLRB1 PLA followed by Lis1/NudE1 immunostaining showed NAGK-dynein complexes were colocalized with Lis1 and NudE1 signals, and PLA for NAGK-Lis1 showed similar signal patterns, suggesting a functional link between NAGK and dynein-Lis1 complex. Subsequently, NAGK-dynein complexes were found in KTs and on nuclear membranes where KTs were marked with CENP-B ICC and nuclear membrane with lamin ICC. Furthermore, knockdown of NAGK by small hairpin (sh) RNA was found to delay cell division. These results indicate that the NAGK-dynein interaction with the involvements of Lis1 and NudE1 plays an important role in prophase nuclear envelope breakdown (NEB) and metaphase MT-KT attachment during eukaryotic cell division.