• BMB Reports
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• v.52 no.6
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• pp.397-402
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• 2019

Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptide of the MERS-CoV S protein and CpG-DNA encapsulated in liposome complex to immunize mice, and produced the monoclonal antibodies 506-2G10G5 and 492-1G10E4E2. The western blotting data showed that both monoclonal antibodies detected the S protein and immunoprecipitated the native form of the S protein. Indirect immunofluorescence and confocal analysis suggested strong reactivity of the antibodies towards the S protein of MERS-CoV virus infected Vero cells. Furthermore, the 506-2G10G5 monoclonal antibody significantly reduced plaque formation in MERS-CoV infected Vero cells compared to normal mouse IgG and 492-1G10E4E2. Thus, we successfully produced a monoclonal antibody directed against the RBD domain of the S protein which could be used in the development of diagnostics and therapeutic applications in the future.

• Journal of Microbiology and Biotechnology
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• v.12 no.5
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• pp.780-786
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• 2002

An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

• Korean Journal of Community Nutrition
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• v.2 no.4
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• pp.523-529
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• 1997

This study was conducted to obtain accurate data on the intake, digestibility and nitrogen balance of protein in Korean college women. Subjects were 8 female college students, aged from 21 to 23, and maintained their menu and life patterns regular during a 4- week study. The same amount of diet that the subjects had consumed, and feces and urine were collected and measured to extract their nitrogen content by Kjeldahl method. From this data, apparent digestibility and the body nitrogen balance were estimated by determing daily protein intake and excretion. The daily protein intake was 56.9$\pm$1.4g and daily fecal protein loss was 6.3$\pm$0.2g. The apparent digestibility of protein was 89.6$\pm$0.7$\%$. The daily nitrogen intake measured by Kjeldahl method was 9.43$\pm$0.2g. The urinary nitrogen excretion was 7.64$\pm$0.23g and fecal nitrogen excretion was 1.02$\pm$0.03g. The nitrogen balance indicated a positive balance of 0.45$\pm$0.18g. (Korean J Community Nutrition 2(4) : 523-529, 1997)

• Food Science and Biotechnology
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• v.18 no.4
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• pp.889-894
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• 2009

Barley proteins are expected to have unique functional properties due to their high content of alcohol soluble protein, hordein. Since the barley proteins obtained by conventional isoelectric precipitation method cannot represent hordein fraction, barley proteins were fractionated to albumin, globulin, glutelin, and hordein with respect to extraction solvents. Functional properties and film-forming properties of solubility-fractionated barley proteins were investigated to explore their potential for human food ingredient and industrial usage. The 100 g of total barley protein comprised 5 g albumin, 23 g globulin, 45 g glutelin, and 27 g hordein. Water-binding capacities of barley protein isolates ranged from 140-183 mL water/100 g solid. Hordein showed the highest oil absorption capacity (136 mL oil/100 g), and glutelin showed the highest gelation property among the fractionated proteins. In general, the barley protein fractions formed brittle and weak films as indicated by low tensile strength (TS) and percent elongation at break (E) values. The salt-soluble globulin fraction produced film with the lowest TS value. Although films made from glutelin and hordein were dark-colored and had lower E values, they could be used as excellent barriers against water transmission.

• Biomolecules & Therapeutics
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• v.27 no.6
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• pp.514-521
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• 2019

G protein-coupled receptors (GPCRs) are membrane receptors whose agonist-induced dynamic conformational changes trigger heterotrimeric G protein activation, followed by GRK-mediated phosphorylation and arrestin-mediated desensitization. Cytosolic regions of GPCRs have been studied extensively because they are direct contact sites with G proteins, GRKs, and arrestins. Among various cytosolic regions, the role of helix 8 is least understood, although a few studies have suggested that it is involved in G protein activation, receptor localization, and/or internalization. In the present study, we investigated the role of helix 8 in dopamine receptor signaling focusing on dopamine D1 receptor (D1R) and dopamine D2 receptor (D2R). D1R couples exclusively to Gs, whereas D2R couples exclusively to Gi. Bioinformatic analysis implied that the sequences of helix 8 may affect GPCR-G protein coupling selectivity; therefore, we evaluated if swapping helix 8 between D1R and D2R changed G protein selectivity. Our results suggest that helix 8 is not involved in D1R-Gs or D2R-Gi coupling selectivity. Instead, we observed that D1R with D2R helix 8 or D1R with an increased number of hydrophobic residues in helix 8 relative to wild-type showed diminished ${\beta}$-arrestin-mediated desensitization, resulting in increased Gs signaling.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2000.04a
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• pp.17-19
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• 2000

Heterotrimeric G Proteins transduce ligand binding to a wide variety of seven transmembrane cell surface receptors into intracellular signals. The currently accepted model for the activation of G protein suggests that ligand-activated receptor accelerates GDP-GTP exchange reactions on the ${\alpha}$ subunit of the heterotrimeric G protein. At least seventeen distinct isoforms of the G${\alpha}$ subunit protein have been identified in mammalian organisms. Among them, the G${\alpha}$q family consists of five members whose ${\alpha}$ subunits show different expression patterns. G${\alpha}$q and G${\alpha}$11 seem to be almost ubiquitously expressed, whereas G${\alpha}$14 is predominantly expressed in spleen, lung, kidney and testis. G${\alpha}$16 and its murine counterpart G${\alpha}$15 are expressed in hematopoietic cells and has been shown to couple a wide variety of receptors to phosphoinositide-specific phospholipase C activity. Beta-isoforms of phospholipase C were shown to be activated by all members of G${\alpha}$q family, i.e., G${\alpha}$q, G${\alpha}$11, G${\alpha}$l4 and G${\alpha}$16 subunits either in reconstitution system. or in experiments using cDNA transfection with intact Cos-7 cells.

• Journal of Nutrition and Health
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• v.24 no.2
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• pp.124-131
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• 1991

This study was to examine the effects of dietary protein levels and the sources of protein(animal and plant) on calcium and phosphorus metabo lism in 10 healthy Korean female adults. The 26­d day study consisted of a 6-day adaptation period and lO-day moderate protein(109 N, 550mg Ca) a and IO-day high protein( 14g N, 570mg Ca) pe­r riod. During the experimental period, the subjects w were divided into two groups, either consuming a animal protein diet(75 % animal protein) or plant protein diet(75 % plant protein). Calcium(300 mg) was supplemented to two subjects of each d dict group for the last 4 days. Feces, urine and diet were analyzed nitrogen. calcium and phos­p phorus. The apparent absorption of calcium was significantly increased as the protein intake was inc­r reased from 60g to 90g in animal protein diet. A Average calcium absorption rate was 30% and 46 % from high and moderate levels of animal p protein diet Urinary calcium excretion was not affected by t the amount of the protein intake in both animal and plant protein diet. There was significantly higher urinary calcium excretion(I34mg) in high I level of animal protein diet than that( 83mg) in h high level of plant protein diet. Calcium balance was improved as the protein intake increased and c calcium was supplemented. Phosphorus absorp­t tion was more efficient in the high animal diet(77.81 %) than in the high plant diet(55-65%). The overall results indicate that an increase of protein and calcium supplement in moderate pro­ttein intake can improve calcium balance due to the increase of calcium absorption.

• Journal of Nutrition and Health
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• v.36 no.9
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• pp.942-949
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• 2003

This study was longitudinally conducted to evaluate the intakes of protein, Ca, Mg and P of exclusively breast-fed infants compared with the Recommended Dietary Allowances (RDA) for Korean infants. Twenty Korean lactating women and their infants during the first 3 months of lactation in Incheon area were participated. Protein, Ca and Mg, and P contents in the milk were determined using semimicro Kjeldahl (N ${\times}$ 6.38) , atomic absorption spectrophotometer and colorimeter, respectively, and also the milk consumption of the infants was measured by the test-weighing method. Protein contents of the milk were 1.96, 1.63, 1.51, 1.25 and 1.16 g/100 ml, and protein intakes of the breast-fed infants were 9.00, 9.85, 9.17, 8.97 and 7.76 g/day at 7, 15, 30, 60 and 90 days postpartum. The average protein intake per body weight of the breast-fed infants was 1.84 g/kg/day. The average intakes of Ca, Mg, P were 172.1 mg/day, 15.2 mg/day and 91.4 mg/day, respectively, and the average Ca/P ratio was 1.91. There was positive correlation between protein and Ca, protein and p, and Ca and P contents while negative correlation between Mg and P, The body weight of breast-fed infants increased normally from 3.6 $\pm$ 0.41 g at birth to three month during lactation. It is suggested that the breast-fed infants in Incheon area consume almost adequately protein, Ca and P from the milk compared with RDA for Korean infants.

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.258-263
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• 1990

A cadmium-binding protein was purified the cell-free extract of extreme cadmium tolerant Hansenula anomala B-7. The molecular weight was determined to be approximately 33, 000 and was composed two kinds of subunits having a molecular weight of 18, 000 and 14, 000, respectively. The extinction coefficient of the cadmium-binding protein was calculated to be 19.58. The amount of cadmium in the cadmium-binding protein was $9.26{\mu}{\textrm{g}}$ per $100{\mu}{\textrm{g}}$ of protein. A total of 14 amino acids were detected in the cadmium-binding protein, including aspartic acid, glycine and alanine that were present in a high quantity, but proline, valine and methionine were not found. The purified cadmium-binding protein contained a high quantity of cysteine and cadmium, and therefore this protein showed clearly the characteristics of metallothionein.

• Journal of Applied Biological Chemistry
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• v.44 no.3
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• pp.125-130
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• 2001

A putative protein encoded by Arabidopsis AtMTN3 gene, a homologue of Medicago truncatula MTN3, consists of 285 amino acid residues, and has a predicted molecular mass of 31.5 kDa and a calculated pI of 9.1. Primary amino acid sequence analyses have revealed that the protein contains seven putative transmembrane regions with N-terminus oriented to the outside of the membrane. The AtMTN3 protein shows overall 16.4% of amino acid identity with the rat GALR3 protein, known to be a G-protein-coupled receptor. The gene is present as a single copy in the Arabidopsis genome, and expressed in aerial parts but not in roots of Arabidopsis. Therefore, AtMTN3 appears not to be specifically involved in Rhizobium-induced nodule development, as was predicted for the MTN3 gene. These proteins possibly mediate signal transmission through G-protein-coupled pathways during general interactions between plants and symbiotic or pathogenic microbes.