• Animal Bioscience
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• 제35권1호
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• pp.96-104
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• 2022

Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons. Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P_2b, P₂₁, and P₂₂) was measured at 3 h and 9 h. Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p<0.05). The phosphorylation levels of 4 bands showed negative associations with immobilized water (P₂₁) and positive association with free water (P₂₂). Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.

• 한국식품과학회지
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• 제20권1호
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• pp.34-39
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• 1988

여러가지 육가공품(肉加工品)중 특정 단백질 원료의 첨가여부를 판정하여 변조식품(變造食品)을 검출하는 한 방법으로서, 각종 육류단백질, non-meat protein, 어육(魚肉)가공품을 대상으로 disc SDS-Poly acrylamide gel electrophoresis의 사용 가능성을 실험하였다. Total protein fraction에 대한 전기영동 결과 복잡하고 많은 band를 보여 각 시료에 고유한 특성을 찾아보기 어려웠다. Low salt-soluble protein fraction에서는 total protein fraction 에서 보다 band 수가 상당히 감소함을 보였고 각 단백질 원료에 대하여 보다 고유한 band pattern을 나타내었다. Acetone-insoluble protein fraction 에서는 non-meat protein의 경우 육류단백질과 상당히 다른 경향을 나타내었고. 소세지 원료의 가열처리에 의하여 단백질의 band수와 양이 감소하였다. 따라서 적당한 단백질 추출조건(抽出條件)을 설정하여 전기영동을 실시하면, 특정(特定) 단백질을 첨가한 변조식품의 검출이 가능해질 것으로 생각된다.

• Reproductive and Developmental Biology
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• 제31권4호
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• pp.253-260
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• 2007

To examine the differential protein expression pattern in the 11.5 day post-coitus (dpc) and 18.5 dpc placenta of mouse, we have used the global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. The differential protein patterns of 3 placentae at the 11.5 dpc and 18.5 dpc from nature mating mice were analyzed. Proteins within isoelectric point range of $3.0{\sim}10.0$, separately were analyzed in 2DE with 3 replications of each sample. A total of approximately 1,600 spots were detected in placental 2-D gel stained with Coomassie-blue. In the comparison of 11.5 dpc and 18.5 dpc placentae, a total of 108 spots were identified as differentially expressed proteins, of which 51 spots were up-regulated proteins such as alpha-fetoprotein, mKIAA0635 protein and transferrin, annexin A5, while 48 spots were down-regulated proteins such as Pre-B-cell colony-enhancing factor l(PBEF), aldolase 1, A isoform, while 4 spots were 11.5 dpc specific proteins such as chaperonin and Acidic ribosomal phosphoprotein P0, while 3 spots were 18.5 dpc specific proteins such as aldo-keto reductase family 1, member B7 and CAST1/ERC2 splicing variant-1. Most identified proteins in this analysis appeared to be related with catabolism, cell growth, metabolism and regulation. Our results revealed composite profiles of key proteins involved in mouse placenta during pregnancy.

• 분석과학
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• 제8권4호
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• pp.575-582
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• 1995

A new analytical procedure using a serial dual electrode detector was developed for the analysis of amino acids and proteins. Bromine was generated at the upstream electrode and detected by the downstream electrode. The presence of amino acids and proteins was shown to lower the downstream current but with no apparent effect on the upstream current. This indirect mode of detection can be applied to the determination of amino acids and proteins which are electrochemically inactive or too large to be accessible to the electrode surface for electron exchange. The method is shown capable to determine various amino acids (cystine, tyrosine, lysine, tryptophan, glycine, methionine and arginine) and proteins (cytochrome c, hemoglobin, HAS, a-Amylase, Conalbumin I, Catalase and Myglobin) with linear working range for amino acids between $10^{-6}$ to $10^{-3}M$ and total proteins between $10^{-7}$ to $10^{-3}M$. The method has been applied for the analysis of amino acids and total protein in food using Flow Injection Analysis with results obtained comparable to those using the traditional analytical procedure. Use of pulse generation technique was shown to produce a more stable flow injection analysis peaks for repetitive determination than the use of conventional constant current method which showed increase of the background current after determination over 200 minutes. The pulse method was found to give stable baseline even after 400 minutes. Thus, the method is shown able to provide a suitable analytical procedure for automatic analysis of amino acids and proteins in food by flow injection analysis.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1519-1528
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• 2017

Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

• 한국초지조사료학회지
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• 제42권3호
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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.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.129-129
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• 2017

The first key point to the successful pollination and fertilization in plants is the pollen pistil interaction, referring to the cellular and molecular levels, which mainly play active roles in limiting gene flow among maize populations and between maize and teosinte. This study was carried out to identify proteins and investigate the mechanism of gametophytic factors using protein analysis. W22 (ga1); which didn't carry a gametophytic factor and W22 (Ga1), a near iso-genic line were used for the proteome investigation. SDS-PAGE was executed to investigate proteins in the pollen and pistil of W22 (ga1) and W22 (Ga1). A total of 44 differentially expressed proteins were identified in the pollen and pistil on SDS-PAGE using LTQ-FTICR MS. Among the 44 proteins, a total of 24 proteins were identified in the pollen of W22 (ga1) and W22 (Ga1) whereas 20 differentially expressed proteins were identified from the pistil of W22 (Ga) and W22 (Ga1). However, in pollen, 2 proteins were identified only in the W22 (ga1) and 12 proteins only in the W22 (Ga1) whereas 10 proteins were confirmed from the both of W22 (ga1) and W22 (Ga1). In contrary, 10 proteins were appeared only in the pistil of W22 (ga1) and 7 proteins from W22 (Ga1) while 3 proteins confirmed in the both of W22 (ga1) and W22 (Ga1). Moreover, the identified proteins were generally involved in hydrolase activity, nucleic acid binding and nucleotide binding. These results help to reveal the mechanism of gametophytic factors and provide a valuable clue for the pollen and pistil research in maize. In addition, it might provide a comprehensive insight on the proteins that were involved in the regulation of pollen-pistil interaction.

• 약학회지
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• 제55권3호
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• pp.203-212
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• 2011

Adipocytes have been known to secrete a number of important proteins called adipokines with roles in energy metabolism, reproduction, cardiovascular function and immunity. In this study we have attempted to identify intensively secretory proteins from 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated into mature adipocytes and then the cells were left in serum-free medium. The supernatant was filtrated and dialyzed. Lyophilized secretome was fractionated by two different methods, 1-D SDS PAGE and RP-FPLC. The tryptic peptides from the gel slices and the FPLC fractions were analyzed by nanoLC/ESI-MS/MS. We identified a total of 303 identical proteins from two methods, 251 proteins from 1-D gel and 184 proteins from RP-FPLC. 86 of them were listed as a secretory protein Finally, we identified many known or unknown secreted proteins existed in the low level including adiponectin, angiotensinogen, bone morphogenetic protein-1 (BMP-1), macrophage migration inhibitory factor (MIF), insulin like growth factor-II (IGF-II), interleukin-6 (IL-6), follistatin-related protein-1, minecan, and resistin. The existence of some of secreted proteins has been confirmed in RNA level. This proteomic experiment is useful for the intensive screening of secretory proteins in many kinds of other cells.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.131-131
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• 2017

Though the Platycodon grandiflorum, has a broad range of pharmacologic properties, but the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two-dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}2-fold$) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, the frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). Taken together, the protein profile may provide insight clues for better understanding the characteristics of proteins and its metabolic activities in various explants of this essential medicinal plant P. grandiflorum.

• 한국작물학회지
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• 제60권1호
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• pp.97-106
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• 2015

Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}$ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.