• Reproductive and Developmental Biology
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• 제37권4호
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• pp.247-253
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• 2013

Here, we present an approach of blood plasma proteome profiling and their comparisons between the young and the adult pigs as prerequisite for the identification of bio-markers related to the health conditions, growth performance and meat quality. To profile the proteome in porcine plasma, blood samples were collected from 19 young piglets and 20 adult male barrows and the plasma was retrieved. Then, protein profiling was initiated using one and two-dimensional electrophoresis. Proteins were spotted and then identified by MALDI-TOF-TOF and LC-MS-MS. In the results, more than thirty-six and twenty eight protein spots were selected in young piglets and adult pigs, respectively and twenty three proteins were identified. The proteome profile images were compared between those ones using Image Master Version 7.0. The image of expressed proteome showed that most of proteins from plasma of young piglet separated clearly and concentrated in 2DE display compared to ones from adult. Image analysis in detail was carried out to look for the specific proteins related to age progression. It demonstrated that the characteristics of proteome expression could be distinct to their age stages. Further investigations needed to proceed to understand the age dependent change of protein conformation and biological meaning of those differences in proteome expression between young and mature adult pigs.

• The Korean Journal of Physiology
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• 제25권2호
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• pp.115-123
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• 1991

Molecular association of glucose transporters with the other proteins in the plasma membrane was assessed by gel electrophoresis and immunoblot techniques. Approximately $31.5{\pm}5.1%$ of GLUT-4, $64.8{\pm}2.7%$ of clathrin, 48.7% of total protein in the plasma membrane (PM) were found insoluble upon extraction with 1% Tx-100. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the Tx-100 insoluble PM fraction contained about 4 major polypeptides with apparent molecular weight of above 200, 100-120, 80 and 30-35 KDa that were readily removed upon wash with a high pH buffer which is known to remove clathrin and 0.5 M Tris-buffer which is known to remove assembly proteins (AP). Immunoblotting of GLUT4 and clathrin against specific antibodies showed that GLUT-4 and clathrin were co-solubilized up to 84.6% and 82.7% respectively by wash with a high pH buffer and 1% Tx-100. When the membrane was pre-washed with a high pH buffer and 0.5 M Tris solution, GLUT4 and clathrin were not solubilized further suggesting that GLUT4 molecules are in molecular association with clathrin, AP and/or other extrinsic membrane proteins in plasma membrane and the formation of clathrin-coated structures might be involved in insulin stimulated glucose transporter translocation mechanism.

• 대한의생명과학회지
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• 제4권1호
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• pp.57-63
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• 1998

본 연구는 돼지 지방세포 원형질막 단백질에 대한 항체를 면양에서 생산하고 생산된 항체의 역가 및 조직특이성을 조사하기 위하여 실시되었다. 지방세포, 뇌, 심장, 신장, 간장 및 비장으로부터 원형질막 단백질을 추출하였으며, 그중 지방세포로부터 분리한 원형질막 단백질을 면양(체중 40kg)에 3주 간격으로 3회 면역 접종시켰다. 면역접종 전, 3차 면역접종 후 10일 (AS-1), 12일 (AS-2)및 14일 (AS-3)째에 각각 면양의 경정맥으로부터 혈액을 채취하여 혈청을 분리하였다. 항체의 역가 및 기타 조직과의 교차반응성은 enzyme-linked immunosorbent assay (ELISA)로 측정하였다. 면양에서 생산된 돼지 지방세포 원형질막 단백질에 대한 항혈청은 지방세포 원형질막 단백질과 강한 항원-항체 반응을 나타내었다. 항혈청의 교차반응성을 조사한 결과, 기타 조직의 원형질막 단백질과는 매우 미약한 반응을 나타낸 반면 지방세포 원형질막 단백질과는 강한 반응을 나타내었다. 이러한 항혈청의 지방세포 원형질막 단백질과의 조직특이적인 반응은 anti-sheep immunoglobulin G-horseradish peroxidase conjugate를 2차 항체로 이용한 immunoblot에 의해서도 재확인되었다. 이상의 결과, 면양으로부터 생산된 돼지 지방세포 원형질막 단백질에 대한 항체는 높은 역가를 지니고 있었으며, 지방세포 원형질막 단백질에 특이적으로 작용함을 알 수 있었다.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2001년도 제41차 추계학술대회
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• pp.57-62
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• 2001

One of recent advances of mammalian fertilization is the understanding of the molecular basis of fertilization. Several proteins localized in sperm nucleus or on sperm surface are necessary for the fertilization process. Protamines, sperm nuclear proteins, are required for normal sperm function that leads to fertilization. Fertilin and cyritestin are sperm surface proteins and essential for sperm-egg binding. Fertilin is also required for sperm transport in the female reproductive tracts. Metalloproteses on sperm plasma membrane are found to play a role in sperm-egg fusion. The functional analysis of these proteins provides a new insight into the molecular mechanisms underlying mammalian fertilization and male fertility.

• Mass Spectrometry Letters
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• 제9권2호
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• pp.51-55
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• 2018

Capture of non-glycoproteins during lectin affinity chromatography is frequently observed, although it would seem to be anomalous. In actuality, lectin affinity chromatography works at post-translational modification (PTM) sites on a glycoprotein which is not involved in protein-protein interactions (PPIs). In this study, serial affinity column set (SACS) using lectins followed by proteomics methods was used to identify PPI mechanisms of captured proteins in human plasma. MetaCore, STRING, Ingenuity Pathway Analysis (IPA), and IntAct were individually used to elucidate the interactions of the identified abundant proteins and to obtain the corresponding interaction maps. The abundant non-glycoproteins were captured with the binding to the selected glycoproteins. Therefore, depletion process in sample pretreatment for abundant protein removal should be considered with more caution because it may lose precious disease-related low abundant proteins through PPIs of the removed abundant proteins in human plasma during the depletion process in biomarker discovery. Glycoproteins bearing specific glycans are frequently associated with cancer and can be specifically isolated by lectin affinity chromatography. Therefore, SACS using Lycopersicon esculentum lectin (LEL) can also be used to study disease interactomes.

• 약학회지
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• 제22권1호
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• pp.42-50
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• 1978

The plasma proteins of fresh water fishes have been fractionated by disc electrophoresis in acrylamide gels utilized as an electrophoretic supporting medium. The species of fishes examined in this experiment were Anguilla japonica, Misgurnus mizolepis, Parasilurus asotus, Siniperca scherzeri, Pelteobagrus fulvidra, Carassius carassius, Cyprinus carpio, and Hemibarbus labeo, obtained in the Han River. Disc electrophoresis was performed as described by Ornstein and Davis. Gels and buffer solution were prepared by the method developed by W.J.Kim. The separation gels were 7% acrylamide gel. The fractionation of plasma proteins showed 13 bands in Anguilla japonica, 10 in Misgurnus mizolepis, 15 in Parasilurus asotus, 12 in Siniperca scherzeri, 11 in Pelteobagrus fulvidra, 13 in Carassius carassius, 9 in Cyprinus carpio, and 13 in Hemibarbus labeo. The patterns of plasma protein on the each species of fishes were different in the number of bands, ratio of contents, relative mobilities, and forms of fractionation.

• Journal of Animal Science and Technology
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• 제47권1호
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• pp.19-28
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• 2005

The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2015년도 제49회 하계 정기학술대회 초록집
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• pp.66.1-66.1
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• 2015

Plasma medicine is an upcoming research area that has attracted the scientists to explore more deeply the utility of plasma. So, apart from the treating biomaterials and tissues with plasma, we have studied the effect of plasma with different feeding gases on modification of biomolecules. Additionally, we have checked the action of nanosecond pulsed plasma on the biomolecules. We have checked the plasma action on proteins ((Hemoglobin (Hb) Myoglobin (Mb) and lysoenzyme), calf thymus DNA and amino acids. The structural changes or structural modification of proteins and DNA have been studied using circular dichroism (CD), dynamic light scattering (DLS), gel electrophoresis, protein oxidation test, UV-vis spectroscopy and 1D NMR, while Liquid Chromatograph/Capillary Electrophoresis-Mass Spectrometer(LC/CE-MS) based qualitative bio-analysis have been used to study the modification of amino acids. We have also shown the effect of NaCl and ionic liquid on the formation of OH radicals using electron spin resonance and fluorescence techinques.

• 한국동물학회지
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• 제19권1호
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• pp.33-42
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• 1976

The blood proteins of ten reptilian species were studied by cellulose acetate elctrophoresis. Three members examined of the genus Agkistrodon have unusually similar patterns in plasma protein, hemoglobin, lactate dehydrogenase and malate dehydrogenase. On the basis of their electrophoretic patterns, it was concluded that Agkistrodon blomhoffii brevicaudus was closely related to the others. In general the plasma protein patterns reflect species specificity. Under the conditions employed, all snakes had a single hemoglobin band except Dinodon rufozonatum rufozonatum which showing two component patterns. Two members of the Chelonia showed four bands of hemoglobin. The zymograms indicated a distinct divergence in blood proteins of the Squamata from those of the Chelonia. The results reflected superficially the established phylogenies of these groups.

• 한국미생물·생명공학회지
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• 제51권2호
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• pp.167-173
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• 2023

Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.