• Korean Journal of Food Science and Technology
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• v.3 no.2
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• pp.101-104
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• 1971

The dye-binding method based on the reaction of acidic orange-G dye with basic groups of protein molecule was investigated to observe its applicability to the determination of protein, basic amino acid and lysine contents in brown rice of several high-protein rice lines. The protein content of rice samples ranged from 7.91 to 10.53% and from 8.93 to 11.96% in terms of wet and dry bases, respectively. The correlation between dye-binding absorbance and protein content in terms of both dry and wet bases was highly significant; their correlation coefficients being $-0.955^{**}\;and\;-0.975^{**}$, respectively. The correlation of dye-binding absorbance lysine and basic amino acids were highly significant and their correlation coefficients were similar. Dye-binding absorbance-lysine showed a lower correlation than dye-binding absorbance-protein but a higher correlation than protein-lysine.

• KSBB Journal
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• v.30 no.6
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• pp.296-301
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• 2015

Eggshell-based biocomposites have become attractive due to their exquisite nanostructure and biological properties, which are mainly composed of highly organized calcium carbonate crystals controlled by organic macromolecules such as proteins and polysaccharides. Here, we designed the recombinant fusion protein of a putative eggshell matrix protein named as GG1234 and a fluorescent reporter protein of DsRed. The protein was successfully over-expressed in E. coli and purified by Ni-NTA affinity chromatography. In vitro calcium carbonate crystallization was conducted in the presence of the fusion protein, and morphological change was investigated. The protein inhibited the calcite growth in vitro, and spherical calcium carbonate micro-particles with the diameter of about $20-30{\mu}m$ were obtained. We expect that this study would be helpful for better understanding of eggshell-based biomineralization.

• Journal of the Korean Magnetic Resonance Society
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• v.21 no.1
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• pp.13-19
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• 2017

Human sterol ${\Delta}8-{\Delta}7$ isomerase, commonly known as emopamil binding protein (EBP), is an essential protein in the cholesterol-synthetic pathway, and mutations of this protein are critically associated with human diseases such as Conradi-Hunermann-Happle or male EBP disorder with neurological defects syndrome. Due to such a clinical importance, EBP has been intensively investigated and some important features have been reported. EBP is a tetra-spanning membrane protein, of which $2^{nd}$, $3^{rd}$, and $4^{th}$ membrane-spanning ${\alpha}$ helices play an important role in its enzymatic function. However, detailed structural feature at atomic resolution has not yet been elucidated, due to characteristic difficulties in dealing with membrane protein. Here, we over-expressed EBP using Escherichia coli and performed detergent screening to find suitable membrane mimetics for structural studies of the protein by NMR. As results, DPC and LMPG could be evaluated as the most favorable detergents to acquire promising NMR spectra for structural study of EBP.

• Animal cells and systems
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• v.2 no.3
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• pp.355-359
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• 1998

Among the proteins secreted from Lactobacillus acidophilus KCTC 3151, a 36 kDA and 24 kDa protein, whose amounts were relatively abundant, were purified and their N-terminal amino acid sequences determined. The N-terminal amino acid sequence of 36 kDa protein exhibited high homology with thymidine phosphorylase and glyceraldehyde-3-phosphate dehydrogenase. The N-terminal amino acid sequence of the 24 kDa protein did not show significant homology with proteins in Protein Data Base nor Gene Bank. Nucleotide sequence of the gene encoding 36 kDa protein indicates that the protein possesses the domains for a-helical, phosphate binding and pyrimidine binding sites, which are also shown in thymidine phosphorylases. Also, the protein contains conserved domains of dehydrogenase II and III. However, the activity of thymidine phosphorylase or glyceraldehyde-3-puospnate dehydrogenase could not be detected in the purified fractions of the 36 kDa protein.

• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.468-473
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• 1996

The genome of tomato spotted wilt virus (TSWV) is composed of three RNA segments, S, M, and L RNA and the 5.0 kb M RNA encodes two glycoproteins Gl, G2 and NSm protein of unknown function. In an effort to investigate the function of the NSm protein, antibody was raised against NSm fusion protein overexpressed in Escherichia coli. This antibody was used to detect the NSm protein by using western blot analysis and electron microscopic observation after immunogold labelling. For the cloning of the NSm gene, total RNA extracted from a TSWV infected plant was used for cDNA synthesis and polymerase chain reaction (PCR) instead of going through time-consuming virus purification. A protein band specifically reacting to the NSm antibody was detected from TSWV inoculated plants. The NSm protein was detected in the cell wall fraction and in pellet from low speed centrifugation when the infected plant tissue was fractionated into 4 fractions. In the immuno-electron microscopic observation, gold particles were found around the plasmodesmata of infected plant tissue. These results suggest that the NSm protein of TSWV plays some role in cell-to-cell movement of this virus.

• BMB Reports
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• v.28 no.6
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• pp.504-508
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• 1995

The electrophoretic patterns of plasma membrane surface proteins of normal rat liver cells and rat hepatomas were compared in 10% non-denaturing and 7-15% gradient non-denaturing gel. Chemical carcinogens, 2-Me DAB (2-methyl-4-dimethylaminoazobenzene) and DENA (diethylnitrosamine), were used to induce hepatoma in rats. One protein which disappeared in hepatoma was identified in normal rat liver by non-denaturing gel electrophoresis. Rabbit antisera were raised against this specific protein, and the protein was purified by Sephacryl S-200 column and immunoaffinity chromatography using the purified antibody. The purified protein showed two bands of molecular weights approximately 50 $kD_{\alpha}$ and 52 $kD_{\alpha}$ by SDS-polyacrylamide gel electrophoresis, which reacted specifically with the antibody. However only one band was observed in non-denaturing gel and also in isoelectric focusing with a pI value of 6.6. This study showed the existence of an unique protein on the plasma membrane surface of normal rat liver cells which disappeared in rat hepatomas induced by chemical carcinogens.

• Journal of Nutrition and Health
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• v.6 no.2
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• pp.1-16
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• 1973

Some effects of dietary conditions on the metabolism of nucleic acid and protein in organs of the Albino Rats have been studied. The young rats to be examined were fed on the control diet and the diets deprived of one component among protein, carbohydrate, and lipid, such as protein free diet (PF: 432 kcal/100g) carbohydrate free diet (CF: 432kcal/100g), and lipid free diet (LF: 392kcal/100g) for three, seven, and fifteen days, respectively. The contents of DNA and RNA in the liver and the brain, and also those of protein-nitrogen(PN) and nonprotein-nitrogen (NPN) in the live, the brain, and the serum have been measured. The results are as followe: 1. The contents of DNA per gram of liver were increased by feeding on protein free diet. It is concluded that the critical factor for the result is not the increase in the rates of DNA syntheses, but the decrease in the turnover rates of DNA. 2. The metabolism of DNA in the liver showed the normal status by feeding on carbohydrate free diet. On the other hand, the rates of DNA syntheses were increased by feeding on lipid free diet. 3. The rates of DNA syntheses in the brain were decreased by feeding on the unbalanced diet, such as protein free, carbohydrete free, and lipid free diet. 4. In the liver and the brain, the rates of DNA syntheses were decreased by feeding on protein free diet. But the rates showed the normal status by feeding on the carbohydrate free diet, and also showed the similar metabolism to that in the case of the control group by feeding on lipid free diet. 5. In the liver, the rates of protein syntheses were decreased, whereas the contents of nonprotein-nitrogen were increased by feeding on protein free diet. 6. In the liver and the brain, the protein syntheses showed the more increasing rates than the rates in the case of the control diet by feeding on lipid free diet. 7. In the serum, the contents of protein did not change in a short period, also the insufficient feeling on protein was examined. It is clear that in the liver the rates of protein syntheses are decreased and the rates of protein catabolism are increased, since the rates of nucleic acid syntheses are decreased by feeding on the protein free diet. On the other hand, it is considered that in the brain the turnover rates of protein does not have correlation with the rates of nucleic acid syntheses, also these are decreased by feeding on protein free diet. And also it is believed that the phenomena of homeostasis for carrying the normal metabolism of nucleic acid and protein in the liver and the brain are operated in a short period as possible, by feeding on carbohydrate free and lipid free diets.

• Archives of Pharmacal Research
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• v.21 no.6
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• pp.712-717
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• 1998

The complete nucleotide sequence of the cDNA clone encoding rat skeletal muscle myosin- binding protein H (MyBP-H) was determined and amino acid sequence was deduced from the nucleotide sequence (GenBank accession number AF077338). The full-length cDNA of 1782 base pairs(bp) contains a single open reading frame of 1454 bp encoding a rat MyBP-H protein of the predicted molecular mass 52.7kDa and includes the common consensus 1CA__TG' protein binding motif. The cDNA sequence of rat MyBP-H show 92%, 84% and 41% homology with those of mouse, human and chicken, respectively. The protein contains tandem internal motifs array (-FN III-Ig C2-FN III- Ig C2-) in the C-terminal region which resembles to the immunoglobulin superfamily C2 and fibronectin type III motifs. The amino acid sequence of the C-terminal Ig C2 was highly conserved among MyBPs family and other thick filament binding proteins, suggesting that the C-terminal Ig C2 might play an important role in its function. All proteins belonging to MyBP-H member contains `RKPS` sequence which is assumed to be cAMP- and cGMP-dependent protein kinase A phosphorylation site. Computer analysis of the primary sequence of rat MyBP-H predicted 11 protein kinase C (PKC)phosphorylation site, 7 casein kinase II (CK2) phosphorylation site and 4N-myristoylation site.

• Animal cells and systems
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• v.5 no.1
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• pp.85-90
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• 2001

Yolk platelets, one of the main food stores in the embryonic development, are composed of proteins. However, little is known about the identity of proteins utilized at certain stages of embryogenesis. In this study, we followed the fates of embryonic storage proteins by using an anti-polyubiquitin monoclonal antibody (mAB) as a probe. The mAb recognized the major storage proteins of Drosophila, Xenopus and chicken eggs. In the Drosophila embryo, the mAb-reactive 45-kDa protein was not used until stage 11 but was used up at stage 16 when the embryo completed segmentation. In the Xenopus embryo, the mAb-reactive 111 kDa protein was mostly utilized between stages 42 and 45 implying that the protein might be an energy source used just prior to feeding on food. By N-terminal sequencing the storage protein of Xenopus embryo was identified as a lipovitellin 1. This study confirms that storage proteins are used almost simultaneously at certain stages of embryogenesis and that vitellogenin 1 is the last storage protein in Xenopus embryogenesis.

• IMMUNE NETWORK
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• v.2 no.3
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• pp.137-141
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• 2002

Among the members of the inhibitor of apoptosis (IAP) protein family, only Livin and survivin have been reported to have variant forms. We have found a variant form of c-IAP2 through the interaction with the X protein of HBV using the yeast two-hybrid system. In contrast to the wild-type c-IAP2, the variant form has two stretches of sequence in the RING domain that are repeated in the C-terminus that would disrupt the RING domain. We demonstrate that the variant form has an inhibitory effect on TNF-mediated $NF-{\kappa}B$ activation unlike the wild-type c-IAP2, which increases TNFmediated $NF-{\kappa}B$ activation. These results suggest that this variant form has different activities from the wild-type and the RING domain may be involved in the regulation of TNF-induced $NF-{\kappa}B$ activation.