• Journal of Life Science
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• v.33 no.11
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• pp.868-875
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• 2023

Kinesin-1 is a motor protein identified as the first member of the kinesin superfamily (KIF), which plays a role in intracellular cargo transport by acting as microtubule-dependent motor proteins within cells. Kinesin-1 consists of two heavy chains (KHCs, also known as KIF5s) and two light chains (KLCs). The 93 amino acids in the carboxyl (C)-terminal tail region of KIF5A are not homologous to the C-terminal tail region of KIF5B or the C-terminal tail region of KIF5C. In this study, we used a yeast two-hybrid screen to identify the binding proteins that interacted with the C-terminal region of KIF5A. We found an association between KIF5A and CUE domain containing 2 (CUEDC2), which is proposed to function as an adaptor protein involved in ubiquitination pathways and protein trafficking. CUEDC2 bound to the C-terminal region of KIF5A and did not interact with KIF5B (the motor of kinesin-1), KIF3A (the motor of kinesin-2), or kinesin light chain 1 (KLC1). KIF5A specifically bound to the C-terminal region of CUEDC2. Furthermore, KIF5A did not interact with another isoform: CUEDC1. In addition, glutathione S-transferase (GST) pull-downs showed that KIF5A directly bound GST-CUEDC2 but did not interact with GST-CUEDC1 and GST alone. When myc-KIF5A and EGFP-CUEDC2 were co-expressed in HEK-293T cells, CUEDC2 co-immunoprecipitated with kinesin-1, and myc-KIF5A and FLAG-CUEDC2 colocalized in the cells. These results suggest that in intracellular cargo transport by kinesin-1, CUEDC2 serves as an adaptor protein connecting kinesin-1 and cargo by binding to KIF5A.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.282-290
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• 1997

Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1125-1132
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• 2013

In the kimchi manufacturing process, the starter is cultured on a large-scale and needs to be supplied at a low price to kimchi factories. However, current high costs associated with the culture of lactic acid bacteria for the starter, have led to rising kimchi prices. To solve this problem, the development of a new medium for culturing lactic acid bacteria was studied. The base materials of a this novel medium consisted of Chinese cabbage extract, a carbon source, a nitrogen source, and inorganic salts. The optimal composition of this medium was determined to be 30% Chinese cabbage extract, 2% maltose, 0.25% yeast extract, and $2{\times}$ salt stock (2% sodium acetate trihydrate, 0.8% disodium hydrogen phosphate, 0.8% sodium citrate, 0.8% ammonium sulfate, 0.04% magnesium sulfate, 0.02% manganese sulfate). The newly developed medium was named MFL (medium for lactic acid bacteria). After culture for 24 hr at $30^{\circ}C$, the CFU/mL of Leuconostoc (Leuc.) citreum GR1 in MRS and MFL was $3.41{\times}10^9$ and $7.49{\times}10^9$, respectively. The number of cells in the MFL medium was 2.2 times higher than their number in the MRS media. In a scale-up process using this optimized medium, the fermentation conditions for Leuc. citreum GR1 were tested in a 2 L working volume using a 5 L jar fermentor at $30^{\circ}C$. At an impeller speed of 50 rpm (without pH control), the viable cell count was $8.60{\times}10^9$ CFU/mL. From studies on pH-stat control fermentation, the optimal pH and regulating agent was determined to be 6.8 and NaOH, respectively. At an impeller speed of 50 rpm with pH control, the viable cell count was $11.42{\times}10^9(1.14{\times}10^{10})$ CFU/mL after cultivation for 20 hr - a value was 3.34 times higher than that obtained using the MRS media in biomass production. This MFL media is expected to have economic advantages for the cultivation of Leuc. citreum GR1 as a starter for kimchi production.

• Journal of Life Science
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• v.24 no.12
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• pp.1276-1283
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• 2014

Cell adhesion molecules determine the cell-cell binding and the interactions between cells and extracellular signals. Cell-cell junctional complexes, which maintain the structural integrity of tissues, consist of more than 50 proteins including multi-PDZ domain protein 1 (MUPP1). MUPP1 contains 13 postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains and serves a scaffolding function for transmembrane proteins and cytoskeletal proteins or signaling proteins, but the mechanism how MUPP1 links and stabilizes the juxtamembrane proteins has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and cell adhesion molecule 1 (Cadm1, also known as SynCAM1, Necl-2, or TSLC1). Cadm1 bound to the second PDZ domain of MUPP1. The carboxyl (C)-terminal end of Cadm1 has a type II PDZ-association motif (-Y-F-I) which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. MUPP1 also bound to the C-terminal cytoplasmic tail region of other Cadm family members (Cadm2, Cadm3, and Cadm4). In addition, these protein-protein interactions were observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-MUPP1 antibody co-immunoprecipitated Cadm1 and Cadm4 with MUPP1 from mouse brain extracts. These results suggest that MUPP1 could mediate interaction between Cadms and cytoskeletal proteins.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.5
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• pp.671-676
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• 2008

Forty-eight Naemi lambs (avg. BW 31.7 kg) were transported by truck for a distance of 1,450 km from Al-Jouf to Riyadh, Saudi Arabia. On arrival day, the lambs were randomly allocated to four groups receiving diets supplemented with 0.0, 0.3, 0.6 and 0.9 ppm organic chromium (Cr). Each group consisted of four separately housed replicates of three lambs each. The animals were fed ad libitum on a grower diet for 84 days. Blood samples were obtained shortly before transportation, upon arrival and at weekly intervals thereafter from all lambs for analysis of plasma and serum. Plasma glucose and serum cortisol, total protein, albumin, urea-N and total cholesterol concentrations were determined. A cursory clinical examination of the lambs, along with rectal temperature, was undertaken at different intervals during the experiment. The lambs were inoculated each with 2 ml i.v. chicken red blood cells (CRBC) on days 0, 21, and 42. Serum total, IgG and IgM antibody titers were determined at weekly intervals post-immunization. An in vivo intradermal hypersensitivity test was carried out on 6 lambs from each group on days 10 and 70. Transportation of the lambs resulted in a significant (p<0.001) elevation of serum cortisol, total protein and albumin levels, as well as increased plasma glucose concentration, with corresponding decrease in total cholesterol, while blood urea-N remained largely unchanged. These constituents returned to normal levels during subsequent weeks, with no significant differences in their concentrations being observed between the Cr-supplemented groups and controls. Rise in rectal temperature after transportation was reduced to a greater extent (p<0.05) in Cr-supplemented versus control lambs. Total, IgG and IgM antibody titers against CRBC rose significantly (p<0.05) during immunizations in all groups, with significantly and linearly higher (p<0.05) total and IgG titers in Cr-supplemented versus control lambs. By contrast, no significant effect due to Cr supplementation was recorded in IgG titers, which increased equally in Cr-fed and control groups. Skin thickness in response to intradermal inoculation of phytohaemagglutinin (PHA) was also significantly (p<0.01) increased as a result of Cr supplementation. These results indicate that dietary Cr supplementation might be useful during stress especially for enhancing immune responses in transport-stressed lambs.

• Journal of Life Science
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• v.19 no.4
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• pp.429-435
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• 2009

Oxidative stress is a consequence of an imbalance of the defense system against cellular damage generated by reactive oxygen species (ROSs) such as superoxide anions (menadione; MD). Most organisms have evolved a variety of defense systems to protect cells from adverse conditions. In order to evaluate stress tolerance against oxidative stress generating MD, comparative analyses of antioxidant capacity, or free radical scavenger ability, were performed between S. cerevisiae KNU5377 (KNU5377) and three wild-type S. cerevisiae strains. In a medium containing 0.4 mM MD, the KNU5377 strain showed higher cell viability and antioxidant ability, and contained higher levels of trehalose, superoxide dismutase, thioredoxin system, glucose-6-phosphate dehydrogenase, and some heat shock proteins. The KNU5377 strain also produced a lower level of oxidative stress biomarker than the other three yeast strains. These results indicate that S. cerevisiae KNU5377 has a higher level of tolerance to oxidative stress due to the increased expression of cell rescue proteins and molecules, thus alleviating cellular damage more efficiently than other S. cerevisiae strains.

• Molecules and Cells
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• v.39 no.5
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• pp.426-438
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• 2016

Plant disease resistance occurs as a hypersensitive response (HR) at the site of attempted pathogen invasion. This specific event is initiated in response to recognition of pathogen-associated molecular pattern (PAMP) and subsequent PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). Both PTI and ETI mechanisms are tightly connected with reactive oxygen species (ROS) production and disease resistance that involves distinct biphasic ROS production as one of its pivotal plant immune responses. This unique oxidative burst is strongly dependent on the resistant cultivars because a monophasic ROS burst is a hallmark of the susceptible cultivars. However, the cause of the differential ROS burst remains unknown. In the study here, we revealed the plausible underlying mechanism of the differential ROS burst through functional understanding of the Magnaporthe oryzae (M. oryzae) AVR effector, AVR-Pii. We performed yeast two-hybrid (Y2H) screening using AVR-Pii as bait and isolated rice NADP-malic enzyme2 (Os-NADP-ME2) as the rice target protein. To our surprise, deletion of the rice Os-NADP-ME2 gene in a resistant rice cultivar disrupted innate immunity against the rice blast fungus. Malic enzyme activity and inhibition studies demonstrated that AVR-Pii proteins specifically inhibit in vitro NADP-ME activity. Overall, we demonstrate that rice blast fungus, M. oryzae attenuates the host ROS burst via AVR-Pii-mediated inhibition of Os-NADP-ME2, which is indispensable in ROS metabolism for the innate immunity of rice. This characterization of the regulation of the host oxidative burst will help to elucidate how the products of AVR genes function associated with virulence of the pathogen.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.287-294
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• 2007

Effect of combined use of anti-microbial materials, such as ethanol, mustard and chitosan, on the quality of low salted kochujang was investigated during fermentation at $20^{\circ}C$ for 12 weeks. Viable cells of yeast increased remarkably during fermentation, but increasing ratio was significantly low in ethanol-mustard added kochujang. Activity of ${\beta}-amylase$ was high in anti-microbial material added kochujang, whereas ${\alpha}-amylase$ and protease activities were low in those groups. Water activity decreased during fermentation with being low in the control kochujang prepared with normal-salt without anti-microbial materials. Hunter L-, a- and b-values of kochujang increased during fermentation, and the degree of increase in total color difference $({\Delta}E)$ was low in ethanol added kochujang. Titratable acidity of kochujang was decreased in anti-microbial materials added group at late aging period, and oxidation-reduction potential was low in the control kochujang. Total sugar and reducing sugar contents of kochujang were high in ethanol-mustard added kochujang. Ethanol contents of kochujang increased at late aging period, with high values in ethanol-chitosan added kochujang. Amino nitrogen content increased during middle of fermentation, and ammonia nitrogen content of kochujang decreased in ethanol-mustard-chitosan added group during fermentation. After 12 weeks fermentation, sensory results showed that ethanol or ethanol-mustard added kochujang were the highest in color and flavor with the highest overall acceptability.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.4
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• pp.273-278
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• 2015

The purpose of this study was to evaluate the in-vitro efficacy of PDT using red light emitting diode (LED) with Radachlorin for biofilm inhibition of clinical Candida albicans isolates. The suspensions containing C. albicans at $9{\times}10^8CFU/mL$ were prepared on yeast nitrogen base containing 5% glucose. The biofilm formation was grown for 3 h after seeding suspensions each 100 ul on a 96-well plate and then supernatant was discarded. Each well was treated with $0.39{\mu}g/mL$ from $50{\mu}g/mL$ concentrations of Radachlorin on adherent biofilm. After a 30-minute incubation, light was irradiated for 30, 60, or 90 minutes using the following light source of wavelength 630 nm LED, at energy densities of 14, 29, and $43J/cm^2$. Afterwards, all supernatant was removed and dried. Adherent cells were stained with safranin O and dried. The cell viability was measured using a microplate reader at 490 nm. Also, a fluorescent signal on C. albicans was observed by saturation of a photosensitizer. In conclusion, a significant inhibition of 72.5% was observed to C. albicans on biofilm at the Radachlorin dose of $50{\mu}g/mL$ with 630 nm LED. The Photosensitizer (Radachlorin) was adequate at 30 minuttes for C. albicans. Overall, the results showed that inhibition of biofilm formation was Radachlorine dose-dependent. The results suggest that PDT, using Radachlorin with 630 nm LED, is able to decrease biofilm formation of C. albicans.

• Korean Journal of Food Science and Technology
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• v.37 no.3
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• pp.449-455
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• 2005

To reduce saft content of kochujang, various combinations of sub-materials such as ethanol mustard and chitosan were added to kochujang, and their effects on microbial characteristics, enzyme activities, and physicochemical characteristics of kochujang were investigated after 12 weeks of fermentation. Activities of ${\beta}$-amylase and pretense were low in ethanol-mustard-chitosan-added kochujang, whereas no significant difference was observed in ${\alpha$-amylase activity among all groups. Number of viable yeast cells decreased remarkably in mustard-added kochujang during late aging period, and anaerobic bacterial counts decreased in sub-material-added groups. Consistency of kochujang increased by addition of sub-materials, and oxidation-reduction potential was low in chitosan-added group. Mustard-chitosan-added kochujang showed lowest increase in total color difference(${\Dalta}E$) and decrease in water activity. PH of kochujang wns highest in mustard-chitosan-added kochujang, resulting in significantly increased titratable acidity. Addition of sub-material increased reducing sugar contents of kochujang, whereas ethanol production was significantly repressed in mustard-chitosan-added kochujang. Amino nitrogen content was Highest in mustard-chitosan-added kochujang during late aging period, whereas ammonia nitrogen content was lower in ethanol-mustard-added kochujang. Results of sensory evaluation indicated ethanol-mustard-added kochujang was more acceptable than other groups in taste and overall acceptability.