• The Korean Journal of Zoology
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• v.26 no.3
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• pp.181-192
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• 1983

Cytosol protein patterns of two strains of A. proteus, tD and xD strain, were compared by two dimensional gel electrophoresis. Among the 200 major polypeptides that could be stained by silver stain method, tD strain contained a cell specific protein whose molecular weight was 45,000 dalton, pI 5.9. On the other hand, the cytosol and the symbiotic vesicles of xD strain contained a symbiosis specific protein (M.W. 29,000; pI 5.5). The fate of the symbiosis specific protein depended on the presence of symbiotic bacteria in the experiment of high temperature effect and of experimental infection. The significance of these results is discussed in relation to their function in organismic association on the basis of the previous findings.

• BMB Reports
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• v.28 no.6
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• pp.477-483
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• 1995

The recombinant human interferon alpha 2a ($rhIFN-{\alpha}2a$), expressed in Saccharomyces cerevtsiae, was purified from insoluble aggregates. The inclusion body of $rhIFN-{\alpha}$ was solubilized by guanidine salt in the presence of disulfide reducing agent. The refolding of denatured $rhIFN-{\alpha}2a$ was achieved by simple dilution. The authentic interferon alpha, which has two correctly matched disulfide bonds, was seperated from incompletely oxidized $IFN-{\alpha}$ and dimeric $IFN-{\alpha}$ by use of a CM-Sepharose column, followed by size exclusion columns at two different pH conditions. The purified protein has been subjected to detailed physicochemical characterization including sequence determination. Unlike other $rhIFN-{\alpha}2a$ from E. coli reported, the $rhIFN-{\alpha}2a$ from S. cerevisiae has no methionine residue at its N-terminus originating from the start codon, ATG. The pI of the protein was determined to be 6.05 with a single band in the pI gel, which demonstrated that the purified $rhIFN-{\alpha}$ was homogeneous. The structural study using circular dichroism showed that the protein retains its three dimensional structure in the wide range of pH conditions between pH 3 and 9, and only minor strucural deformation was observed at pH 1.0.

• Journal of The Korean Society of Grassland and Forage Science
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• v.42 no.3
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• pp.137-145
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• 2022

Aluminum (Al) is one of the major factors adversely affects crop growth and productivity in acidic soils. In this study, the effect of Al on plants in soil was investigated by comparing the protein expression profiles of alfalfa roots exposed to Al stress treatment. Two-week-old alfalfa seedlings were exposed to Al stress treatment at pH 4.0. Total protein was extracted from alfalfa root tissue and analyzed by two-dimensional gel electrophoresis combined with MALDI-TOF/TOF mass spectrometry. A total of 45 proteins differentially expressed in Al stress-treated alfalfa root tissues were identified, of which 28 were up-regulated and 17 were down-regulated. Of the differentially expressed proteins, 7 representative proteins were further confirmed for transcript accumulation by RT-PCR analysis. The identified proteins were involved in several functional categories including disease/defense (24%), energy (22%), protein destination (9%), metabolism (7%), transcription (5%), secondary metabolism (4%), and ambiguous classification (29%). The identification of key candidate genes induced by Al in alfalfa roots will be useful to elucidate the molecular mechanisms of Al stress tolerance in alfalfa plants.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.92-102
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• 2006

Abundant proteins often cause problems in proteome study. Glutelin family proteins (hereafter referred to glutelin) are present in rice proteome sample as over-whelming constituents with very high abundance. In order to increase the number of identified proteins in rice proteome study, we developed a newly improved method for sample preparation through the removal of glutelin. When the protein samples from rice seed were extracted by the conventional trichloroacetic acid (TCA) extraction method, glutelin accounts for about 60% of total rice seed proteins in SDS gels. Using our new water extraction method, glutelin consists of only about 10% of total proteins. After analyzing on a two-dimensional gel electrophoresis (2-DE), 937 protein spots were detected using the conventional TCA extraction method. On the other hand, 1240 proteins could be seen using the new water extraction method. The selectivity for non-glutelin and less abundant protein by the water extraction method was also confirmed by ESI-Q/TOF mass spectrometry analysis. Thus, the new water extraction method developed here can be efficiently used to study the proteome analysis of rice storage seed.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.9
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• pp.1239-1246
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• 2016

The breeding of yaks is highly seasonal, there are many crucial proteins involved in the reproduction control program, especially in follicular development. In order to isolate differential proteins between mature and immature follicular fluid (FF) of yak, the FF from yak follicles with different sizes were sampled respectively, and two-dimensional gel electrophoresis (2-DE) of the proteins was carried out. After silver staining, the Image Master 2D platinum software was used for protein analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was performed for differential protein identification. The expression level of transferrin and enolase superfamily member 1 (ENOSF1) was determined by Western blotting for verification analysis. The results showed that 2-DE obtained an electrophoresis map of proteins from mature and immature yak FF with high resolution and repeatability. A comparison of protein profiles identified 12 differently expressed proteins, out of which 10 of them were upregulated while 2 were downregulated. Western blotting showed that the expression of transferrin and ENOSF1 was enhanced with follicular development. Both the obtained protein profiles and the differently expressed proteins identified in this study provided experimental data related to follicular development during yak breeding seasons. This study also laid the foundation for understanding the microenvironment during oocyte development.

• Korean Journal of Soil Science and Fertilizer
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• v.20 no.3
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• pp.275-283
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• 1987

In order to find out the effectiveness of nitrogen fixation in rhizobia-legume symbiotic relationship, ecological and physiological characteristics of indigenous rhizobia distributed in Korean soils, that is, dissimilatory nitrate reduction patterns of indigenous soybean rhizobia isolated from four different soils, and differences in one-and two-dimensional polyacrylamide gel electrophoretic pattern of proteins among the each subgroups of Bradyrhizobium japonicum and Rhizobium fredii, were investigated. The results were summarized as follows: 1. The indigenous soybean rhizobia isolated from four different soils could be classified into 4 groups depending on growth rate and dissimilatory nitrate reduction pattern, that is, $S_1$ (slow-grower; Bradyrhizobium japonicum and nitrate denitrifier), $S_2$ (slow-grower; Bradyrhizobium japonicum and nitrate respirer), $F_1$ (fast-grower; Rhizobium fredii and denitrifier), and $F_2$ (fast-grower; Rhizobium fredii and nitrate respirer). 2. The population ratio of fast- to slow-growing R. japonicum was 39% to 61%, and the ratio of denitrifier to nitrate respirer was 31% to 69% and 89% to 11% in fast and slow-grower, respectively. Some differences were observed between fast- and slow-growing R. japonicum but no significant difference was observed between denitrifier and nitrate respirer within same growth type by one and two dimensional SDS-polyacrylamide gel electrophoretic patterns.

• The Korean Journal of Zoology
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• v.29 no.2
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• pp.86-106
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• 1986

For the comparative study of protein patterns of the sera of pregnant women, the protein in sera of normal male subjects, non-pregnant women, pregnant women and women delivered of children were analyzed by using the methods of SDS/polyacrylamide gel electrophoresis, two-dimensional electrophoresis and amino acid analysis. The results were as follows: 1. When the protein patterns of sera in normal male ranging from 10, 000 to 110, 000 daltons were compared to non-pregnant women by SDS/polyacrylamide gel electrohporesis, their protein patterns were same each other numerically but bands 3(22, 000 dalton) and 6(39, 000) were less in male than in non-pregnant women quantitatively. When the protein patterns in the pregnant women in which serum were collected two week intervals were compared with non-pregnant women, there was increased or decreased in several bands quantitavely. The protein patterns of sera in pregnant women were compared with those of non-pregnant women; band 3(22, 000) showed similar patterns each other until the 16th week but the quantity of protein was decreased continously from the 18th week to the third trimester of pregnancy. Contary, bands 4(24, 000), 9(69, 000), 10(70, 000), 12(80, 000), 14(86, 000), 15(91, 000) and 16(94, 000) were gradually increased in quantity from the begining of gestation, and band 7(51, 000) was increased until the 32th week of gestation only but somewhat decreased after this time. The quantities of bands 12(80, 000), 15(91, 000) and 16(94, 000) were relatively increased when the protein patterns of delivered women were compared with those of the third trimester of pregnancy. Women who were dilivered female children showed more increase in bands 4(24, 000), 7(51, 000) and 10(70, 000)than one who were delivered male chilren. 2. When the protein patterns of sera in normal males were compared with those of nonpregnant women by two-dimensional electrophoresis, three spots of spot a group were not appeared in the males and the spot c group in the males was less than in the non-pregnant women. In the pregnant women, albumin was significantly decreased during the 10-12 week of gestation but recovered after these times. And spot f(70, 000) was decreased in the 10th week of gestation but increased from this time. 3. Glutamic acid, lysine, aspartic acid, leucine and valine in pregnant women were large in quantity while methionine, isoleucine and glycine were small in quantity by amino acid analysis. The total amino acids were increased remarkably in the second trimester of pregnancy but began to decrease in the third trimester of pregnancy. As mentioned, this present paper deat with that proteins which consist of maternal serum were increased with specific period in pregnancy and that the change of characteristic protein patterns were identified in the serum protein of each trimester in the pregnant women. And furthermore, the study should be preformed for the sex-identification of a fetus in the early pregnancy.

• Reproductive and Developmental Biology
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• v.32 no.4
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• pp.255-261
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• 2008

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

• Journal of Applied Biological Chemistry
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• v.54 no.3
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• pp.178-183
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• 2011

The stem woods of Lindera obtusiloba Blume were extracted in 80% aqueous methanol and the concentrated extract was partitioned with ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc and n-BuOH fractions, five compounds were isolated through the repeated silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies. On the basis of spectroscopic data including mass spectrometry, IR, $^1H$-NMR, $^{13}C$-NMR, distortionless enhancement by polarization transfer, and two-dimensional-NMR gradient correlated spectroscopy (gCOSY), gradient heteronuclear single quantum correlation (gHSQC), gradient heteronuclear multiple bonding connectivity (gHMBC), the chemical structures of the compounds were determined as asarinin (1), (+)-catechin (2), (-)-epicatechin (3), hyperin (4), and nudiposide (5). Compounds 1 and 5 were isolated for the first time from the stem wood of L. obtusiloba Blume.

• Proceedings of the Korean Vacuum Society Conference
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• 2012.02a
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• pp.246-246
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• 2012

Smart catalyst design though novel catalyst preparation methods can improve catalytic activity of transition metals on reducible oxide supports such as titania by enhancement of metal oxide interface effects. In this work, we investigated Pt nanoparticles/titania catalysts under CO oxidation reaction by using novel preparation methods in order to enhance its catalytic activity by optimizing metal oxide interface. Arc plasma deposition (APD) and metal impregnation techniques are employed to achieve Pt metal deposition on titania supports which are prepared by multi-target sputtering and Sol-gel techniques. In order to tailor metal-support interface for catalytic CO oxidation reaction, Pt nanoparticles and thin films are deposited in varying surface coverages on sputtered titania films using APD. To assess the role of oxide support at the interface, APD-Pt is deposited on sputtered and Sol-gel prepared titania films. Lastly, characteristics of APD-Pt process are compared with Pt impregnation technique. Our results show that activity of Pt nanoparticles is improved when supported over Sol-Gel prepared titania than sputtered titania film. It is suggested that this enhanced activity can be partly ascribed to a very rough titania surface with the higher free metal surface area and higher number of sites at the interface between the metal and the support. Also, APD-Pt shows superior catalytic activity under CO oxidation as compared to Pt impregnation on sputtered titania support. XPS results show that bulk oxide is formed on Pt when deposited through impregnation and has higher proportion of oxidized Pt in the form of $Pt^{2+/4+}$ oxidation states than Pt metal. APD-Pt shows, however, mild oxidation with large proportion of active Pt metal. APD-Pt also shows trend of increasing CO oxidation activity with number of shots. The activity continues to increase with surface coverage beyond 100%, thus suggesting a very rough and porous Pt films with higher active surface metal sites due to an increased surface area available for the reactant CO and $O_2$ molecules. The results suggest a novel approach for systematic investigation into metal oxide interface by rational catalysts design which can be extended to other metal-support systems in the future.