• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.307-307
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• 1994

본 실험에서는 유전자 재조합으로 제조한 〔$^{125}$/I〕-labeled human GM-CSF를 사용하여 GM-CSF의 HL-60 cell의 표면에 존재하는 GM-CSF 수용체의 특성을 밝히고 수용체에 대한 binding parameter를 확인하고 Immunex(미국) 사에서 제조한 Prokine(Sargramostim)과 Sigma사에서 구입한 GM-CSF(C-9666)를 표준물질로 하여 (주) Lucky에서 제조한 GM-CSF(LBD-005)의 수용체에 대한 결합율을 측정, 각각 비교하고자 하였다. 한편 LBD-005는 glycosylation된 form과 안된 form의 혼합물이므로 당화의 정도가 수용체에 대한 결합에 미치는 영향을 알아보기 위해 glycosylated form과 혼합물(LBD-005)의 수용체에 대한 결합율을 측정 비교하였다.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2003년도 International Meeting of the Microbiological Society of Korea
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• pp.122-124
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• 2003

Cyclodextrin glucanotransferase (CGTase) and endoxylanase genes of Bacillus sp. were subcloned down-stream of yeast ADH1 promoter and expressed in S. cerevisiae. Most of the CGTase and endoxylanase expressed were detected in the extracellular medium with activity of 0.6 and 7-8 unit/ml, respectively. The recombinant enzymes were secreted as N-linked-glycosylated forms, resulting an enhanced thermal stability. CGTase predominantly produced $\alpha-cyclodextrin$ from starch and endoxylanase produced xylobiose and xylotriose from xylan.

• 분석과학
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• 제8권4호
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• pp.849-854
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• 1995

A series of biologically active phosphopeptides were synthesized and their behavior in tandem mass spectrometry have been investigated. The structure identifications of other unusual peptides such as sulphated, glycosylated, lipoidal, and backbone modified peptides have been carried out. For all tested peptides, the structural modification could be determined directly by measurement of the absolute molecular weight in combination with collision-induced-dissociation in tandem mass spectrometry.

• Childhood Kidney Diseases
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• 제10권1호
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• pp.1-7
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• 2006

Osteopontin (OPN) is a glycosylated phosphoprotein which mediates cell adhesion and migration, and is produced by bone, macrophages, endothelial cells, and epithelial cells. The many regulatory functions of OPN include bone remodeling, tumor invasion, wound repair, and promotion of cell survival. It is produced by renal tubular epithelial cells, and expression is upregulated in glomerulonephritis, hypertension, ischemic acute renal failure, renal ablation, and UUO. In this review, we discuss about osteopontin in general aspect, expression, role on the development and pathologic condition of neonatal kidney.

• 한국식품영양학회지
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• 제22권2호
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• pp.159-165
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• 2009

E. japonica is a well-known medicinal plant in Japan. The leaves of E. japonica were reported to have a hypoglycemic action. However, seeds of E. japonica are discarded and not used. To elucidate for anti-diabetic effects of E. japonica, Type 2 diabetic mice were allocated to control group, E. japonica leaf, and seed extract group. Animals were fed a 2018S Teklad global 18% protein rodent diet. Animals were received daily oral injections of E. japonica leaf or seed extract at a dose of 200 mg/kg body weight for 6 weeks. Body weight, food intake and water intake, and total adipose tissue weight of animals were significantly reduced by feeding of E. japonica leaf extract. All E. japonica extract groups significantly decreased fasting blood glucose, glycosylated hemoglobin levels, size of adipocytes and serum adiponectins. However, they did not have a beneficial effect on the serum triglyceride and cholesterol in the diabetic animals. These results suggest that E. japonica seed and leaf extracts have a antidiabetic effect by controlling of blood glucose and decrease of size of adipocytes in db/db mice and seed extract is more effective in hypoglycemic action than leaf extract.

• 동의생리병리학회지
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• 제27권2호
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• pp.196-201
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• 2013

Abnormal regulation of glucose and impaired lipid metabolism that result from a defective or deficient insulin are the key etiological factor in type 2 diabetes mellitus (T2DM). The our study evaluated the beneficial effect of diet supplementation with Lentinus edodes on hyperglycemia and lipid metabolism in normal and type 2 diabetic rats. The animals were divided into 4 groups: group I(control) rats were fed standard diet (12% of calories as fat); group II (T2DM) rats were fed HFD (40% of calories as fat) for 2 weeks and then injected with STZ (50 mg/kg); group III and group IV rats were continually fed a diet containing 1% and 10% Lentinus edodes for 4 weeks after T2DM induction, respectively. After 4 weeks we determined biochemical parameters such as glucose, insulin concentration, serum total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), and glycosylated hemoglobin (HbA1c) concentration were also measured. There was a significant reduction in serum TC and TG in the Lentinus edodes supplement groups. The Lentinus edodes diet supplementation were found to have a potent lipid metabolism improvement as well as LDL concentration decreased and HDL concentration was increased. Concentrations of blood glucose and HbA1c in the experimental groups II were significantly decreased after 4 weeks compared with the control group. The Lentinus edodes diet supplementation is useful in regulating the glucose level, improves the insulin, HbA1c, serum lipid metabolism in experimental diabetic rats. We suggest that Lentinus edodes supplementation may have the control effects of diabetes mellitus by improving blood glucose control and lipid metabolism.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.383-391
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• 2008

The glycans linked to the insect cell-derived glycoproteins are known to differ from those expressed in mammalian cells, partly because of the low level or lack of glycosyltransferase activities. GnT II, GnT IV, GnT V, and ST3Gal IV, which play important roles in the synthesis of tetraantennarytype complex glycan structures in mammalian cells, were overexpressed in Trichoplusia ni cells by using a baculovirus expression vector. The glycosyltransferases, expressed as a fusion form with the IgG-binding domain, were secreted into the culture media and purified using IgG sepharose resin. The enzyme assay, performed using a pyridylaminated-sugar chain as an acceptor, indicated that the purified glycosyltransferases retained their enzyme activities. Human erythropoietin expressed in T. ni cells (rhEPO) was subjected to in vitro glycosylation by using recombinant glycosyltransferases and was converted into complex-type glycan with terminal sialic acid. The presence of Nacetylglucosamine, galactose, and sialic acid on the rhEPO moiety was detected by a lectin blot analysis, and the addition of galactose and sialic acid to rhEPO was confirmed by autoradiography using $UDP-^{14}C-Gal\;and\;CMP-^{14}C-Sia$ as donors. The in vitro glycosylated rhEPO was injected into mice, and the number of reticulocytes among the ed blood cells was counted using FACS. A significant increase in the number of reticulocytes was not observed in the mice injected with in vitro glycosylated rhEPO as compared with those injected with rhEPO.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권4호
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• pp.234-241
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• 2000

The Saccharomyces cerevisiae PMR1 gene encodes a Ca2+-ATPase localized in the Golgi. We have investigated the effects of PMR1 disruption in S. cerevisiae on the glycosylation and secretion of three heterologous glycoproteins, human ${\alpha}$1-antitrypsin (${\alpha}$1-AT), human antithrombin III (ATHIII), and Aspergillus niger glucose oxidase (GOD). The pmr1 null mutant strain secreted larger amounts of ATHIII and GOD proteins per a unit cell mass than the wild type strain. Despite a lower growth rate of the pmr1 mutant, two-fold higher level of human ATHIII was detected in the culture supernatant from the pmr1 mutant compared to that of the wild-type strain. The pmr1 mutant strain secreted ${\alpha}$1-AT and the GOD proteins mostly as core-glycosylated forms, in contrast to the hyperglycosylated proteins secreted in the wild-type strain. Furthermore, the core-glycosylated forms secreted in the pmr1 mutant migrated slightly faster on SDS-PAGE than those secreted in the mnn9 deletion mutant and the wild type strains. Analysis of the recombinant GOD with anti-${\alpha}$1,3-mannose antibody revealed that GOD secreted in the pmr1 mutant did not have terminal ${\alpha}$1,3-linked mannose unlike those secreted in the mnn9 mutant and the wild type strains. The present results indicate that the pmr1 mutant, with the super-secretion phenotype, is useful as a host system to produce recombinant glycoproteins lacking high-mannose outer chains.

• Journal of Microbiology and Biotechnology
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• 제27권11호
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• pp.1916-1924
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• 2017

In this study, we synthesized a glycosylated derivative of caffeic acid phenethyl ester (CAPE) using the amylosucrase from Deinococcus geothermalis with sucrose as a substrate and examined its solubility, chemical stability, and anti-inflammatory activity. Nuclear magnetic resonance spectroscopy showed that the resulting glycosylated CAPE (G-CAPE) was the new compound caffeic acid phenethyl ester-4-O-${\alpha}-{\small{D}}$-glucopyranoside. G-CAPE was 770 times more soluble than CAPE and highly stable in Dulbecco's modified Eagle's medium and buffered solutions, as estimated by its half-life. The glycosylation of CAPE did not significantly affect its anti-inflammatory activity, which was assessed by examining lipopolysaccharide-induced nitric oxide production and using a nuclear factor erythroid 2-related factor 2 reporter assay. Furthermore, a cellular uptake experiment using high-performance liquid chromatography analysis of the cell-free extracts of RAW 264.7 cells demonstrated that G-CAPE was gradually converted to CAPE within the cells. These results demonstrate that the glycosylation of CAPE increases its bioavailability by helping to protect this vital molecule from chemical or enzymatic oxidation, indicating that G-CAPE is a promising candidate for prodrug therapy.

• Biomolecules & Therapeutics
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• 제14권1호
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• pp.30-35
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• 2006

Tissue-type plasminogen activator (t-PA) is a multidomain serine protease containing two kringle domains, TK1-2. Previously, Pichia-derived recombinant human TK1-2 has been reported as an angiogenesis inhibitor although t-PA plays an important role in endothelial and tumor cell invasion. In this work, in order to improve in vivo efficacy of TK1-2 through elimination of immune reactivity, we mutated wild type TK1-2 into non-glycosylated form (NE-TK1-2) and examined whether it retains anti-angiogenic activity. The plasmid expressing NE-TK1-2 was constructed by replacing $Asn^{l17}\;and\;Asn^{184}$ with glutamic acid residues. After expression in Pichia pastoris, the secreted protein was purified from the culture broth using S-sepharose and UNO S1-FPLC column. The mass spectrum of NE-TK1-2 showed closely neighboring two peaks, 19631.87 and 19,835.44 Da, and it migrated as one band in SDS-PAGE. The patterns of CD-spectra of these two proteins were almost identical. Functionally, purified NE-TK1-2 was shown to inhibit endothelial cell migration in response to bFGF stimulation at the almost same level as wild type TK1-2. Therefore, the results suggest that non-glycosylated NETK1-2 can be developed as an effective anti-angiogenic and anti-tumor agent devoid of immune reactivity.