• Title/Summary/Keyword: protein structures

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Changes in Spectroscopic Characteristics and Pyrene Binding Reactivities of Dissolved Organic Matters By Biodegradation (생분해에 의한 용존 자연유기물질 분광특성 및 Pyrene 결합반응성 변화)

  • Park, Min-Hye;Hur, Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.9
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    • pp.893-899
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    • 2008
  • Changes in spectroscopic characteristics and pyrene binding coefficients of terrestrial dissolved organic matters(DOM) were investigated during microbial incubation. The incubation studies were conducted for 21 days using a leaf litter DOM and a soilderived DOM with an inoculum from a river. The dissolved organic carbon(DOC), the specific UV absorbance(SUVA), the synchronous fluorescence spectra, and the pyrene organic carbon-normalized binding coefficient(K$_{oc}$) of the DOM were measured at the incubation days of 0, 3, 7, 14 and 21. After the 21-day incubation, DOC were reduced to 61% and 51% of the original concentrations of the litter DOM and the soil-derived DOM, respectively. Comparison of the spectroscopic characteristics before and after the incubation revealed that the SUVA, the fulvic-like fluorescence(FLF), the humic-like fluorescence(HLF) of the different DOM were enhanced by the incubation whereas the protein-like fluorescence(PLF) was reduced. This indicates that more aromatic and humic-like compounds were enriched during the biodegradation process while biodegradable and weak carbon structures were depleted. Irrespective of the DOM sources, SUVA values showed a positive relationship with pyrene K$_{oc}$ with a correlation coefficient of 0.97. The FLF and HLF also exhibited good correlations with K$_{oc}$ values although different regression equations were obtained from the different DOM. Our results suggest that the selected spectroscopic characteristics could be good estimation indices for the changes of the binding reactivity of DOM for hydrophobic organic contaminants during biodegradation process.

Purification of Band 3 from the Human Erythrocyte Membrane and its Incorporation into Liposome (사람 적혈구막 Band 3의 정제 및 Liposome으로의 도입)

  • Kim, Jae-Ryong;Kim, Jung-Hye;Lee, Ki-Yung
    • Journal of Yeungnam Medical Science
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    • v.3 no.1
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    • pp.41-48
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    • 1986
  • Band 3, the predominent 95,000 dalton anion transport protein, is the major intrinsic glycoprotein of the human erythrocyte membrane. This anion carrier exists as a dimer and binds the cytoskeletons such as spectrin, ankyrin and actin. And the liposomes are vesicular structures which form spontaneouly upon hydration of phospholipids. These artificial lipid vesicles have been investigated as model of the biological membranes and as a mean of improving the delivery of nucleic acids, drugs, proteins and biological substances to specific target tissues and cells. In this study, we were purified Band 3 from the human erythrocyte membrane(ghost) was prepared by hemolysis of intact human erythrocyte with weak alkali-hypotonic solution. Band 6 was removed from ghost by extracting with solution of an ionic strength of 0.15. Band 3 and Band 4 were solubilized selectively by extracting Band 6-depleted ghosts with Triton X-100 under nondenaturing conditions. Band 3 was then purified from Triton X-100 extract treated with p-chloromercuribenzoate by sucrose density gradient ultracentrifugation. This purified Band 3 was incorporated into liposomes prepared by reverse-phase evaporation. Phosphatidyl L-serine and cholesterol(1 : 1 molar ratio) were dissolved in chloroform and then chloroform was removed by rotatory evaporation under reduced pressure. Band 3 solution without Triton X-100 was introduced into a mixture of lipids and diethylether. Diethylether was subsequently removed by evaporation. This purified Band 3 and its incorporation into liposomes were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

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Molecular Cloning and Nucleotide Sequence Analysis of pyrB Gene Encoding Aspartate Transcarbamylase from Psychrophilic Sporosarcina psychrophilia (저온성균 Sporosarcina psychrophilia로부터 Aspartate Transcarbamylase 유전자의 클로닝 및 염기서열 분석)

  • 성혜리;안원근;김사열
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.312-319
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    • 2002
  • The Sporosarcina psychrophilia pyrB gene, which encodes aspartate transcarbamylase (ATcase), was cloned on Sau3AI restriction endonuclease fragment inserted into pUC19 plasmid vector, S. psychrophilia pyrB gene was expressed in Escherichia coli pyrB mutant for the complementation test. The sequence of 2,606 nucleotides including putative pyrB gene was determined. The region contained one full open reading frame (ORf) and two partial ORFs. The deduced amino acid sequence of the second ORF showed 59% identity with that of Bacillus caldolyticus ATCase. The first and third partial ORFs were closely related to the uracil permease (pyrP) and dihydroorotase (pyrC), respectively. Besides, potential terminator, antiterminator, and anti-antiterminator structures were found in the intergenic region between pyrP and pyrB. These results suggested that S. psychrophilia pyrimidine nucleotide biosynthesis genes are clustered as well as other Bacillus sp. Over-expressed product of pyrB encoding ATCase was purified and analyzed by the SDS-PAGE. The purified PyrB protein turned out to be molecular mass of 27 kDa and showed ATCase activity.

The Isolation and Pyrolysis of the Brown Pigmented Macromolecule from the Cured Leaf Tobacco (잎담배 성분중 갈색고분자 물질의 분리정제 및 열분해에 관한 연구)

  • Chae, Quae;Park, Ji-Chang
    • Journal of the Korean Society of Tobacco Science
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    • v.2 no.1
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    • pp.1-7
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    • 1980
  • Gel filtration column chromatography (Sephadex G-75), dialysis an d Brushite column chromatography were carried out to separate the brown pigmented macromolecule from water extracts of the cured leaf tobaccos. The two distinct macromolecules having different molecular weight were separated by the Sephadex column chromatography. Brushite also separated two different species of macromolecules which might have different electronic structures. According to the enzymatic degradation of protein in Burley and Hicks, chymotrypsin showed the best degradation ratio, ie., 16-30% in Burley and 38-57% in Hicks. Similar effect was observed with pepsin. However, very low effect of degradation was revealed with trypsin. The sample treated with the proteolytic enzymes revealed the disappearance of the first peak and the slight decrease of the 2nd peak height in the separation profile of Sephadex. After dialysis, the brown pigmented macromolecule was pyrolyzed at $300^{\circ}C$ and the strongly fluorescent components not identified before pyrolysis were detected with TLC separation. Absorption spectrum of these fluorescent compounds was monitored in benzene and the absorption maxima at 265nm and 275 nm were obtained. Considering absorption maxima and shape of the spectrum, those fluorescent compounds seem to be PAH derivatives.

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EFFECT OF RHPDGF-BB AND RHBMP-2 ON OSSEOINTEGRATION OF TITANIUM IMPLANTS AT PERIIMPLANT BONE DEFECTS GRAFTED WITH HYDROXYAPATITE: MICRO-CT AND HISTOLOGIC ANALYSIS (Hydroxyapatite를 이식한 임플란트 주위 골결손부에서 rhPDGF-BB와 rhBMP-2가 골내 임플란트 osseointegration에 미치는 영향: Micro-CT 분석과 조직학적 평가)

  • Park, Jee-Hyun;Hwang, Sun-Jung;Kim, Myung-Jin
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.6
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    • pp.461-468
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    • 2009
  • Purpose: Platelet derived growth factor(PDGF)-BB and bone morphogenetic protein(BMP)-2 are well-known representative growth factors. The purposes of this study were to investigate the effect of rhPDGFBB and rhBMP-2 on osseointegration of titanium implants at periimplant bone defects grafted with hydroxyapatite and to evaluate the feasibility of imaging bone structures around screw-type titanium implant with micro-CT. Materials and Methods: The first molar and all premolars in the mandible region of four beagle dogs were extracted. Following a healing period of 4 months, three $8{\times}8{\times}6mm$-sized bony defects were formed and screw-type titanium implants were placed with hydroxyapatite(HA) block and growth factors; Control group, PDGF group and BMP group. Two months post-implantation, the mandible was harvested. Bone volume(BV), bone-to-implant contact(BIC) and bone mineral density(BMD) were analyzed with micro-CT and histology. Results: According to micro-CT analysis, BV and BMD measures of PDGF and BMP group were significantly higher than control group(BV; PDGF group: $p{\fallingdotseq}0.011$, BMP group: $p{\fallingdotseq}0.006$/BMD; PDGF group: $p{\fallingdotseq}0.020$, BMP group: $p{\fallingdotseq}0.011$) and BIC measures of BMP group were significantly higher than PDGF group($p{\fallingdotseq}0.015$). In histologic evaluation, BIC measures of BMP group was significantly higher than PDGF group($p{\fallingdotseq}0.048$). The values of BV in histologic sections were higher than in micro-CT images and the values of BIC in micro-CT images were higher than in histologic sections. Conclusion: The findings of this experimental study indicates that the use of rhPDGF-BB and rhBMP-2 can increase new bone formation in a large bony defect around titanium implant, and rhBMP-2 is more effective than rhPDGF-BB. Micro-CT can be considered useful for assessment as a rapid and nondestructive method for 3-dimensional measurement of bone healing around implants. Further study is necessary, however, to remove metal artifacts around titanium implant and to standardize the method.

Identification and gene expression profiling of chicken Pumilio family, Pum1 and Pum2

  • Lee, Jee-Young;Kim, Duk-Kyung;Zheng, Ying-Hui;Kim, Sun-Young;Kim, Hee-Bal;Lim, Jeong-Mook;Han, Jae-Yong
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2005.11a
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    • pp.64-65
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    • 2005
  • Members of the Pumilio are the RNA binding proteins acting as translational repressors and required for germ cell development and asymmetric division. We identified chicken Pum1 and Pum2 that are similar to mouse and human in highly conserved C-terminal RNA-binding domain and eight tandem repeats. The comparative sequence analysis of Pum1 and Pum2 from fly, chicken, mouse and human shows high degree of evolutionary conservation in the homology of the peptide sequence and the structure of PUM-HD (Pumilio homology domain) with similar spacing between adjacent Pum repeats. Also, structures of chicken Pum1 and Pum2 genes are almost identical to those of mouse and human. We revealed that the expression levels of Pum1 and Pum2 were the highest in hatched female gonad among various embryonic tissues, and Pum2 expressed highly in 12-day and hatched gonad by real-time RT-PCR. These results suggest that Pum1 and Pum2 might have an effect on the development of chicken gonad.

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Molecular analysis of alternative transcripts of equine AXL receptor tyrosine kinase gene

  • Park, Jeong-Woong;Song, Ki-Duk;Kim, Nam Young;Choi, Jae-Young;Hong, Seul A;Oh, Jin Hyeog;Kim, Si Won;Lee, Jeong Hyo;Park, Tae Sub;Kim, Jin-Kyoo;Kim, Jong Geun;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.10
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    • pp.1471-1477
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    • 2017
  • Objective: Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified AXL receptor tyrosine kinase (AXL) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. Methods: We validated two isoforms of AXL transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Results: Both of AXLa and AXLb transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between AXLa and AXLb transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. Conclusion: It is assumed that the expression patterns of AXLa and AXLb transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an $NF-{\kappa}B$ signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.

Viroid-the Smallest Plant Pathogen (바이로이드-가장 작은 식물병원체)

  • Lee Jai Youl
    • Korean Journal Plant Pathology
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    • v.1 no.3
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    • pp.199-206
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    • 1985
  • Viroids are the smallest. well-characterized infectious agents presently known. and so far viroids have been found only in higher plants. The structures of viroid-molecules are single-stranded, covalently closed circular RNA molecules with a range of 240 to 380 nucleotides according to the various viroids. Viroids are remarkable not only as a new category of pathogen, which cause economically important diseases, but also as an excellent model system for biochemical and biophysical investigations because of their small size, relative stability and their self-replication. Four different patato spindle tuber viroid isolates, which express the different symptoms on the same host plant exchange only 2 to 6 nucleotides in the total number of 359 nucleotides, but now the mechanism of viroid pathogenicity is not explained fully. Viroid-melecules are replicated without any special viroid-associated proteins, and during the process of viroid replication oligomeric viroid-associated RNAs are detected at nuclei of viroid infected leaf tissue. The mechanism of viroid replication can now be illustrated according to a possible explanation of rolling-circle system. Although the rapid progress have been made in elucidation of the biochemical and biophysical properties of PSTV and other viroids, the mechanism of viroid replication and pathogenicity is less known and is still a matter of speculation. When these problems can be sufficiently explained, the viroid molecule could play an important role as an available vector in plant genetic engineering.

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Roles of Prostatic Acid Phosphatase in Prostate Cancer (Prostatic acid phosphatase의 전립선 암에서의 역할)

  • Kong, Hoon-Young;Lee, Hak-Jong;Byun, Jong-Hoe
    • Journal of Life Science
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    • v.21 no.6
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    • pp.893-900
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    • 2011
  • Prostatic acid phosphatase (PAP) is one of the widely used biomarkers in the diagnosis of prostate cancer. It was initially identified in 1935 and is the most abundant phosphatase in the human prostate. PAP is a prostate-specific enzyme that is synthesized in prostate epithelial cells. It belongs to the acid phosphatase group that shows enzymatic activity in acidic conditions. PAP is abundant in prostatic fluid and is thought to have a role in fertilization and oligospermia. It also has a potential role in reducing chronic pain. But one of the most apparent functions of PAP is the dephosphorylation of macromolecules such as HER-2 and PI3P that are involved in the ERK1/2 and MAPK pathways, which in turn leads to inhibition of cell growth and tumorigenesis. Currently, clinical trials using PAP DNA vaccine are underway and FDA-approved immunotherapy using PAP is commercially available. Despite these clinically important aspects, molecular mechanisms underlying PAP regulation are not fully understood. The promoter region of PAP was reported to be regulated by NF-${\kappa}B$, TNF-${\alpha}$, IL-1, androgen and androgen receptors. Here, the features of PAP gene and protein structures together with the function, regulation and roles of PAP in prostate cancer are discussed.

Inhibitory Effects of Naeso-san on Pacemaker Potentials in Interstitial Cells of Cajal of Murine Small Intestine (생쥐 소장 카할세포의 내향성 향도잡이 전압에 미치는 내소산의 억제효과에 관한 연구)

  • Hong, Noo Ri;Ahn, Tae Seok;Park, Hyun Soo;Chae, Han;Kwon, Young Kyu;Kim, Byung Joo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.6
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    • pp.630-635
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    • 2014
  • The purpose of this study was to investigate the effects of Naeso-san in interstitial cells of Cajal (ICCs) in murine small intestine. First, we isolated ICCs from murine small intestine. After that, we cultured these cells for 1 days. The patch-clamp technique was applied on ICCs that formed network-like structures in culture (1 days). Spontaneous rhythms were routinely recorded from cultured ICCs under current-clamp conditions, and the ICCs within networks displayed more robust electrical rhythms (pacemaker potentials). To understand the relationship between Naeso-san and pacemaker activity in ICCs, we examined the effects of Naeso-san on pacemaker potentials of ICCs. In current clamp mode (I = 0), the addition of Naeso-san (10 mg/ml - 50 mg/ml) decreased the amplitude and frequency of the pacemaker potentials of ICCs in a dose dependent manner. However, these effects were blocked by intracellular $GDP{\beta}S$, a G-protein inhibitor, and glibenclamide, a specific ATP-sensitive K+ channels blocker. Pretreatment with SQ-22536, an adenylate cyclase inhibitor, did not block the Naeso-san induced effects, whereas pretreatment with ODQ, a guanylate cyclase inhibitor, or L-NAME, an inhibitor of nitric oxide (NO) synthase blocked the Naeso-san induced effects. Our findings provide insight into unraveling the modulation of Naeso-san in pacemaker potentials of ICCs and developing therapeutic agents against gastrointestinal motility disorders.