• Bulletin of the Korean Chemical Society
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• v.29 no.2
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• pp.381-388
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• 2008

An X-linked skeletal disorder, SEDT (spondyloepiphyseal dysplasia tarda) is a genetic disease characterized by a disproportionately short trunk and short stature caused by mutations in the SEDL gene. This gene is evolutionarily conserved from yeast to human. The yeast SEDL protein ortholog, Trs20p, has been isolated as a member of a large multi-protein complex called the transport protein particle (TRAPP), which is involved in endoplasmic reticulum (ER)-to-Golgi transport. The interaction between SEDL and partner proteins is important in order to understand the molecular mechanism of SEDL functions. We isolated several SEDL-binding proteins derived from rat cells by an immobilized GST-fusion method. Furthermore, the SEDL-homologue proteins were identified using motif based methods. Common motifs between SEDL-binding proteins and SEDL-homologue proteins were classified into seven types and 78 common motifs were revealed. Sequence similarities were contracted to seven types using phylogenetic trees. In general, types I-III and VI were classified as having the function of acetyl-CoA carboxylase, glycogen phosphorylase, isocitrate dehydrogenase, and enolase, respectively, and type IV was found to be functionally related to the GST protein. Types V and VII were found to contribute to TRAPP vesicle trafficking.

• Molecules and Cells
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• v.37 no.3
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• pp.257-263
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• 2014

A mammalian cell renovates itself by autophagy, a process through which cellular components are recycled to produce energy and maintain homeostasis. Recently, the abundance of gap junction proteins was shown to be regulated by autophagy during starvation conditions, suggesting that transmembrane proteins are also regulated by autophagy. Transient receptor potential vanilloid type 1 (TRPV1), an ion channel localized to the plasma membrane and endoplasmic reticulum (ER), is a sensory transducer that is activated by a wide variety of exogenous and endogenous physical and chemical stimuli. Intriguingly, the abundance of cellular TRPV1 can change dynamically under pathological conditions. However, the mechanisms by which the protein levels of TRPV1 are regulated have not yet been explored. Therefore, we investigated the mechanisms of TRPV1 recycling using HeLa cells constitutively expressing TRPV1. Endogenous TRPV1 was degraded in starvation conditions; this degradation was blocked by chloroquine (CLQ), 3MA, or downregulation of Atg7. Interestingly, a glucocorticoid (cortisol) was capable of inducing autophagy in HeLa cells. Cortisol increased cellular conversion of LC3-I to LC-3II, leading autophagy and resulting in TRPV1 degradation, which was similarly inhibited by treatment with CLQ, 3MA, or downregulation of Atg7. Furthermore, cortisol treatment induced the colocalization of GFP-LC3 with endogenous TRPV1. Cumulatively, these observations provide evidence that degradation of TRPV1 is mediated by autophagy, and that this pathway can be enhanced by cortisol.

• Molecules and Cells
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• v.42 no.4
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• pp.292-300
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• 2019

Immunometabolism, defined as the interaction of metabolic pathways with the immune system, influences the pathogenesis of metabolic diseases. Metformin and carbon monoxide (CO) are two pharmacological agents known to ameliorate metabolic disorders. There are notable similarities and differences in the reported effects of metformin and CO on immunometabolism. Metformin, an anti-diabetes drug, has positive effects on metabolism and can exert anti-inflammatory and anti-cancer effects via adenosine monophosphate-activated protein kinase (AMPK)-dependent and AMPK-independent mechanisms. CO, an endogenous product of heme oxygenase-1 (HO-1), can exert anti-inflammatory and antioxidant effects at low concentration. CO can confer cytoprotection in metabolic disorders and cancer via selective activation of the protein kinase R-like endoplasmic reticulum (ER) kinase (PERK) pathway. Both metformin and CO can induce mitochondrial stress to produce a mild elevation of mitochondrial ROS (mtROS) by distinct mechanisms. Metformin inhibits complex I of the mitochondrial electron transport chain (ETC), while CO inhibits ETC complex IV. Both metformin and CO can differentially induce several protein factors, including fibroblast growth factor 21 (FGF21) and sestrin2 (SESN2), which maintain metabolic homeostasis; nuclear factor erythroid 2-related factor 2 (Nrf2), a master regulator of the antioxidant response; and REDD1, which exhibits an anticancer effect. However, metformin and CO regulate these effects via different pathways. Metformin stimulates p53- and AMPK-dependent pathways whereas CO can selectively trigger the PERK-dependent signaling pathway. Although further studies are needed to identify the mechanistic differences between metformin and CO, pharmacological application of these agents may represent useful strategies to ameliorate metabolic diseases associated with altered immunometabolism.

• Biomolecules & Therapeutics
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• v.30 no.6
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• pp.546-552
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• 2022

Epidermal cell adhesion molecule (EpCAM) is a tumor-associated antigen (TAA), which has been considered as a cancer vaccine candidate. The EpCAM protein fused to the fragment crystallizable region of immunoglobulin G (IgG) tagged with KDEL endoplasmic reticulum (ER) retention signal (EpCAM-FcK) has been successfully expressed in transgenic tobacco (Nicotiana tabacum cv. Xanthi) and purified from the plant leaf. In this study, we investigated the ability of the plant-derived EpCAM-FcK (EpCAM-FcK^P) to elicit an immune response in vivo. The animal group injected with the EpCAM-FcK^P showed a higher differentiated germinal center (GC) B cell population (~9%) compared with the animal group injected with the recombinant rhEpCAM-Fc chimera (EpCAM-Fc^M). The animal group injected with EpCAM-FcK^P (~42%) had more differentiated T follicular helper cells (Tfh) than the animal group injected with EpCAM-Fc^M (~7%). This study demonstrated that the plant-derived EpCAM-FcK fusion antigenic protein induced a humoral immune response in mice.

• Journal of Life Science
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• v.13 no.5
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• pp.596-603
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• 2003

Cells respond to an accumulation of unfolded proteins in the endoplasmic reticulum (ER) by increasing transcription of genes encoding molecular chaperones and folding enzymes. The information is transmitted from the ER lumen to the nucleus by intracellular signaling pathway, called the unfolded protein response (UPR). To obtain genes related to UPR from B. mori, the cDNA library was constructed with mRNA isolated from Bm5 cell lines in which N-glycosylation was inhibited by tunicamycin treatment. From the cDNA library, we selected 40 clones that differentially expressed when cells were treated with tunicamycin. Among these clones, we have isolated ATFC gene showing similarity with Hac1p, encoding a bZIP transcription factor of 5. cerevisiae. Basic-leucine zipper (bZIP) domain in amino acid sequences of ATFC shared homology with yeast Hac1p. Also, ATFC is up-regulated by accumulation of unfolded proteins in the ER through the treatment of ER stress drugs. Therefore we suggest that ATFC represents a major component of the putative transcription factor responsible for the UPR leading to the induction of ER-localized stress proteins.

• Applied Microscopy
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• v.30 no.2
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• pp.141-152
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• 2000

This study aims to demonstrate the effect of squalene (SQ), one of the natural chelator, on the ultrastructural changes in the mouse liver caused by $CdCl_2$. A total of 40 healthy ICR that weighted 30 gm $({\pm}2gm)$ was used for experiment. The experimental group was divided into two groups; group A and B. The group A administrated $CdCl_2$ (4.0 mg/kg) to the intraperitoneal. The group B administrated $CdCl_2$ (4.0 mg/kg) to the intraperitoneal treated with SQ (180 mg/kg, 2 time/day). Each group was observed at 24, 48, 72, 96 hours after injected $CdCl_2$. The results were as follows: 1. Group A Nuclear membrane was observed very irregular at the 24 hours and keep rounded-shape from 48 hours. Mitochondria were observed destruction of inner cavity to the 72 hours and some showed inner cavity destruction at 96 hours. RER (rough endoplasmic reticulum) showed the dilation of inner cavity all the around and reformed typical lamellae from 72 hours. Lysosome were observed in the cytoplasm from 24 hours. From 72 hours, glycogen showed over cytoplasm. 2. Group B Nuclear membrane was observed regular at overall the time. Mitochondria showed normal shapes (round, rod) at overall the time. To 48 hours, inner cavity of rER dilated and destructed lamellae. But from 72 hours, observed typical lamellae of rER Lysosome were observed from 24 hours. And glycogen showed from 24 hours. These results suggest that squalene attenuates the toxic effect of the $CdCl_2$ in the mouse liver.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.3
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• pp.398-407
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• 2020

Objective: The Yip1 domain family (YIPF) proteins were proposed to function in endoplasmic reticulum (ER) to Golgi transport and maintenance of the morphology of the Golgi, which were homologues of yeast Yip1p and Yif1p. YIPF3, the member 3 of YIPF family was a homolog of Yif1p. The aim of present study was to investigate the expression and regulation mechanism of porcine YIPF3. Methods: Quantitative realtime polymerase chain reaction (qPCR) was used to analyze porcine YIPF3 mRNA expression pattern in different tissues and pig kidney epithelial (PK15) cells stimulated by polyinosine-polycytidylic acid (poly [I:C]). Site-directed mutations combined with dual luciferase reporter assays and electrophoretic mobility shift assay (EMSA) were employed to reveal transcription regulation mechanism of porcine YIPF3. Results: Results showed that the mRNA of porcine YIPF3 (pYIPF3) was widely expressed with the highest levels in lymph and lung followed by spleen and liver, while weak in heart and skeletal muscle. Subcellular localization results indicated that it expressed in Golgi apparatus and plasma membranes. Upon stimulation with poly (I:C), the level of this gene was dramatically up-regulated in a time- and concentration-dependent manner. pYIPF3 core promoter region harbored three cis-acting elements which were bound by ETS proto-oncogene 2 (ETS2), zinc finger and BTB domain containing 4 (ZBTB4), and zinc finger and BTB domain containing 14 (ZBTB14), respectively. In which, ETS2 and ZBTB4 both promoted pYIPF3 transcription activity while ZBTB14 inhibited it, and these three transcription factors all played important regulation roles in tumorigenesis and apoptosis. Conclusion: The pYIPF3 mRNA expression was regulated by ETS2, ZBTB4, and ZBTB14, and its higher expression in immune organs might contribute to enhancing ER to Golgi transport of proteins, thus adapting to the immune response.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.221-232
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• 1993

Morphological changes in the uterine and vaginal epithelial cells of the Korean native goats were studied in fifteen primiparous goats slaughtered on the day of parturition and on days 1, 3, 10 and 21 postpartum. 15 uterus and vagina from goats were examined by scanning and transmission electron microscopy. The results obtained in this study were summarized as follows : 1. Transmission electron microscopically, long microvilli which sometimes ramified were found until 10 days postpartum, while short microvilli were found at 21 days. The high electron dense irregular-shaped mitochondria were found in the cytoplasm and the crystalline structure of the mitochondrial matrix was also found from 1 day to 10 days postpartum. Well-developed rough-endoplasmic reticulum (rER) with dilated cisternae which contained the proteins materials was observed at 21 days postpartum. These materials were fused each other and then large granules were found in the free surface of the cytoplasm. A few lipid droplets were generally appeared in the cytoplasm, while numerous droplets were found at 21 days postpartum. A moderate number of ribosomes, a few multivesicular bodies, vesicles, lysosomes and macrophages were found. The globule leucocytes were observed from 0 to 3 days postpartum by transmission electron microscopy. The short microvilli, high electron dense cytoplasm and severe indentation of the nuclear enbelope were found in the vaginal epithelium. Numerouos small vesicles and a few vacuoles were observed in the apical cytoplasmic portion of the epithelium. A few mitochondria were high electron dense and irregular in shape. A moderate amounts of microfilaments, loose intercellular space and dilated rER were also found at 21 days postpartum. 2. Scanning electron microscopically, the folds of the uterine mucosa were generally deep. The long microvilli of the epithelium were found until 3 days postpartum, while short microvili were found at 10 and 21 days postpartum. The distinct intercellular boundary was seen. The apporcine secretory profile of the epithelium observed at between 3 and 10 days postpartum and the cells were somewhat protruded into the lumen. The short microvilli were found on the surface of the protruded cells, while polygonal microridge profile of the epithelium and some dome-shaped epithelium were also observed at 21 days postpartum. The folds of the vaginal mucosa were deep and epithelium was polygonal in shape. The microvilli of the epithelium were long until 3 days postpartum, while they were short at 10 and 21 days. The polygonal epithelium was invaginated into the center of the cell surface until 10 days postpartum. The microridge and dome in shape of the epithelium were found at 10 days postpartum, while the polygonal and exfoliating epithelium were observed at 21 days.

• Applied Microscopy
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• v.27 no.1
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• pp.71-86
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• 1997

The regeneration and differentiation of the cutaneous pigment system in the goldfish, Carassius auratus during the wound healing process were studied with high magnification electron microscope. The cutaneous pigment cells of the normal tissues were composed of three kinds of dermal chromatophores-xanthophores, leucoiphores and melanophores. While xanthophores contain two kinds of pigment granules-pterinosomes and carotenoid vesicles, leucophores and melanophores contain amorphous pigment granules (leucosomes) and oval shaped electron dense melanin pigment granules (melanosomes) respectively. After injury, primary wound healing responses being carried out by migration of epidermal cells and hemocytes spreading over the wound surface at the day of wounding. And at the time of primary wound closure, 5 to 7 days after wounding, rER rich cells-presumably common precursors of dermal chromatophores-immigrated into the wound area. First redifferentiated chromatophores appeared 3 weeks after wounding. Pigment granules of the chromatophores were emerged from the cytoplasmic Golgi complex via rough endoplasmic reticulum. Pinocytotic vesicles which associated with accumulation of pigment material, appeared only at the inner surface of the chromatophores adhering to the rER rich cells, characteristically. The differentiation of each chromatophore in addition to integumental wound repair were accomplished within 4 weeks after wounding at most cases, however the total numbers and densities of these repaired chromatophores still primitive state. Moreover, It has been revealed that complete repair of chromatophores at wounded tissues from burns requirs more than 3 months in normal environment.

• Molecules and Cells
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• v.25 no.3
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• pp.390-396
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• 2008

Calreticulin (CRT) is one of the major $Ca^{2+}$ binding chaperone proteins of the endoplasmic reticulum (ER) and an unusual luminal ER protein. Postnatally elevated expression of CRT leads to impaired development of the cardiac conductive system and may be responsible for the pathology of complete heart block. In this study, the molecular mechanisms that affect $Ca^{2+}$-dependent signal cascades were investigated using CRT-overexpressing cardiomyocytes. In particular, we asked whether calreticulin plays a critical role in the activation of $Ca^{2+}$-dependent apoptosis. In the cells overexpressing CRT, the intracellular calcium concentration was significantly increased and the activity of PKC and level of SECAR2a mRNA were reduced. Phosphorylation of Akt and ERKs decreased compared to control. In addition the activity of the anti-apoptotic factor, Bcl-2, was decreased and the activities of pro-apoptotic factor, Bax, p53 and caspase 8 were increased, leading to a dramatic augmentation of caspase 3 activity. Our results suggest that enhanced CRT expression in mature cardiomyocytes disrupts intracellular calcium regulation, leading to calcium-dependent apoptosis.