• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1708-1713
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• 2002

Endometrial explants obtained from does between days 13 and 21 of pregnancy were cultured in a modified minimum essential medium in the presence of [$^35S$]methionine and [$^3H$]-leucine. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. No marked qualitative changes in patterns of protein production by caprine endometrium between days 13-21 of pregnancy. At least 11 proteins showed consistently a clear spot or a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa). Limited N-terminal sequence analysis of these two isoforms showed that the protein had complete homology with bovine placental and plasma retinol-binding protein (RBP) over the first 20 amino acids. Through use of the antiserum raised against bovine placental RBP, immunoreactive RBP was detected in cultures conditioned by uterine explants prepared at days 13, 15 and 21 of pregnancy. In the present study, proteins synthesized and secreted by caprine endometrium during periattachment period of early pregnancy were characterized. The pregnant endometrium secreted a number of neutral-to-acidic proteins which constituted, in part, the histotroph. A vitamin A-transport protein, RBP, was identified in cultures conditioned by endometrium of days 13-21 of pregnancy. The uterine endometrium is the only source of retinol for embryonic tissues. The uterine RBP appears to transport retinol locally toward embryonic tissues. Secretion of RBP by caprine endometrium of days 13, 15 and 21 of pregnancy suggested that retinol played an important role in conceptus development during periattachment period of early pregnancy.

• Asian-Australasian Journal of Animal Sciences
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• 제16권7호
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• pp.962-967
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• 2003

The purpose of the study were to characterize the proteins secreted by elongating caprine conceptus, to identify a group of low molecular weight proteins as retinol-binding protein (RBP), to identify RBP cell-specific localization in conceptus tissue, and to demonstrate that the conceptuses secreted continuously RBP during the time period maternal recognition of pregnancy. Caprine conceptuses were removed from the uterus between days 16 and 22 of pregnancy, the time period maternal recognition of pregnancy. Isolated conceptuses were cultured in a modified minimum essential medium in the presence of radiolabeled amino acids. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. At least five proteins showed consistently a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa) was identified as RBP by using antiserum against RBP. Presence of RBP in conceptus culture medium and uterine flushings between days 16 and 22 of pregnancy were determined by immunoprecipitation and Western blotting using anti-RBP serum. In immunocytochemical study, strong immunostaining for RBP was localized in trophectoderm and endoderm of conceptus. These results clearly demonstrated that the caprine conceptus was active in protein synthesis as early as day 16 of pregnancy. Secretion of RBP by caprine conceptuses (days 16-22) coincident with the rapid transformation of the conceptus from a spherical blastocyst to a filamentous structure. Production of RBP by the elongating conceptuses may be indicative of an important role for conceptus RBP in the transport, availability and metabolism of retinol during maternal recognition of pregnancy.

• International Journal of Industrial Entomology and Biomaterials
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• 제16권2호
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• pp.37-48
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• 2008

The diapause and non diapause associated proteins of multivoltine silkworm eggs were analysed by two dimensional (2D) gel electrophoresis. The study was made at 0 hr, 24 hrs and 48 hrs after oviposition. A total of four protein spots in diapause eggs at 24 hrs of oviposition and two protein spots in non diapause eggs at 0 hrs of oviposition were observed. All the six protein spots were considered to have association with diapause and non diapause characters. The molecular weight (MW) and isoelectric point (PI) of these 6 protein spots were calculated. The protein spots 1 and 2 observed in 0 hr of non diapause eggs were found to have the MW of 67 and 75 KDa and PI of 8.6 and 8.4 respectively. Similarly the four protein spots observed in diapause egg at 24 hrs of oviposition exhibited MW viz., 15, 17,20 and 25 KDa and PI of 5.3, 5.8, 6.5 and 6.0 respectively. All these 6 identified protein spots were subjected to in-gel digestion and resulted tryptic peptides were analyzed by Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI TOF-MS). Databases searched based on experimentally determined molecular weights of peptides for the determination of the identities of proteins. The identified proteins indicated homology of 34% to 95%. The results indicate that the proteins may playa role in development of diapause and non diapause eggs.

• 한국식품과학회지
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• 제18권5호
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• pp.339-344
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• 1986

Trypsin inhibitor가 trypsin과 안정한 복합체를 형성하고 동시에 일반 단백질은 trypsin에 의하여 가수분해 되는 원리를 이용하여 trypsin inhibitor를 용이하게 검정 할 수 있는 방법을 고안하였다. trypsin으로 가수분해 시킨 대두추출액을 Sephadex G-50을 이용하여 trypsin-trypsin inhibitor 복합체를 분리시킨 후에 SDS 전기 영동으로 trypsin inhibitor를 복합체를 분리시킨 후에 SDS 전기 영동으로 trypsin inhibitor를 검정할 수 있었다. 이들 trypsin inhibitor는 trypsin에 의한 2차 가수분해에서도 가수분해 되지 않았으며, 또한 2차원 전기영동과 DEAE-Sephades A-25크로마토 그래피를 이용하여 trypsin inhibitor가 trypsin과 복합체를 형성하는 능력을 검정함으로써 본 방법의 유효성을 확인하였다. 본 실험 방법으로 대두(Hill 품종)의 trypsin inhibitor를 검정한 결과 7개의 trypsin inhibitor를 찾아 낼 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.895-902
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• 2012

The Korean native chickens (Woorimotdak$^{TM}$, KNC) and commercial broilers (Ross, CB) show obvious differences in meat flavor after cooking. To understand the contribution of protein and peptide for meat flavor, 2-dimensional (2-D) gel electrophoresis and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry was performed. A total of 16 protein spots were differentially expressed in the breast and thigh meat between the two breeds. A total of seven protein spots were represented by different levels between KNC and CB for breast meat. Among them three protein spots (TU39149, TU40162 and TU39598) showed increases in their expressions in KNC while other four protein spots (BU40125, BU40119, BU40029 and BU39904) showed increases in CB. All nine protein spots that were represented by different levels between KNC and CB for thigh meat showed increases in their expression in KNC. Phosphoglucomutase 1 (PGM 1), myosin heavy chain (MyHC), heat shock protein B1 (HSP27), cytochrome c reductase (Enzyme Q), Glyoxylase 1, DNA methyltransferase 3B (DNA MTase 3) were identified as the main protein spots by MALDI-TOF mass spectrometry. These results can provide valuable basic information for understanding the molecular mechanism responsible for breed specific differences in meat quality, especially the meat flavour.

• Journal of Microbiology
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• 제44권4호
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• pp.375-382
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• 2006

From its initial colonization to causation of disease, Streptococcus pneumoniae has evolved strategies to cope with a number of stressful in vivo environmental conditions. In order to analyze a global view of this organism's response to heat shock, we established a 2-D electrophoresis proteome map of the S. pneumoniae D39 soluble proteins under in vitro culture conditions and performed the comparative proteome analysis to a 37 to $42^{\circ}C$ temperature up-shift in S. pneumoniae. When the temperature of an exponentially growing S. pneumoniae D39 culture was raised to $42^{\circ}C$, the expression level of 25 proteins showed changes when compared to the control. Among these 25 proteins, 12 were identified by MALDI-TOF and LC-coupled ESI MS/MS. The identified proteins were shown to be involved in the general stress response, energy metabolism, nucleotide biosynthesis pathways, and purine metabolism. These results provide clues for understanding the mechanism of adaptation to heat shock by S. pneumoniae and may facilitate the assessment of a possible role for these proteins in the physiology and pathogenesis of this pathogen.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.288-295
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• 2015

Salmonella enterica serovar Enteritidis is the predominant agent causing salmonellosis in chickens and other domestic animals. In an attempt to identify antigenic S. Enteritidis outer membrane proteins (OMPs) that may be useful for subunit vaccine development, we established a proteomic map and database of antigenic S. Enteritidis OMPs. In total, 351 and 301 spots respectively from S. Enteritidis strain 270 and strain 350 were detected by two-dimensional gel electrophoresis. Fifty-one antigen-reactive spots were detected by antisera on two-dimensional immunoblots and identified as 12 specific proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. OmpA and DNA starvation/stationary phase protection protein (Dps) were the most abundant proteins among the identified OMPs, comprising 22 and 12 protein species, respectively. Interestingly, we found that the Dps of S. Enteritidis is also antigenic. OmpW was also verified to have high antigenicity. These results show that OmpA, Dps, and possibly OmpW are antigenic proteins. This study provides new insights into our understanding of the immunogenic characteristics of S. Enteritidis OMPs.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.130-130
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• 2017

Sorghum bicolor is considered as copper-tolerant species. The present study was conducted to understand the copper tolerance mechanism in Sorghum seedling roots. Morphological and effects of Cu on other interacting ions were observed prominently in the roots when the plants were subjected to different concentrations (0, 50, and $100{\mu}M$) of $CuSO_4$. However, the morphological characteristics were reduced by Cu stress, and the most significant growth inhibition was observed in plants treated with the highest concentration of $Cu^{2+}$ ions ($100{\mu}M$). In the proteome analysis, high-throughput two-dimensional polyacrylamide gel electrophoresis coupled with MALDI-TOF-TOF mass spectrometry was performed to explore the molecular responses of Cu-induced sorghum seedling roots. In two-dimensional silver-stained gels, a total of 422 differentially expressed proteins (${\geq}1.5-fold$) were identified using Progenesis SameSpot software. A total of 21 protein spots (${\geq}1.5-fold$) from Cu-induced sorghum roots were analyzed by mass spectrometry. Of the 21 differentially expressed protein spots from Cu-induced sorghum roots, a total of 10 proteins were up-regulated, and 11 proteins were down-regulated. The abundance of the most identified protein species from the roots that function in stress response and metabolism was significantly enhanced, while protein species involved in transcription and regulation were severely reduced. The results obtained from the present study may provide insights into the tolerance mechanism of seedling roots in Sorghum.

• 한국작물학회지
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• 제59권3호
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• pp.359-363
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• 2014

Soybean seed is a good source of plant protein in human consumables such as baby formula and protein concentrate. The seeds contain an abundance of storage proteins, namely ${\beta}$-conglycin and glycinin that account for ~ 70-80% of the total seed protein content. Proteome profiling has been proved to be an efficient way that can help us to investigate the seed storage proteins. In the present study, the seeds were removed from the pods and the cotylendonary tissues were separated from the testa for proteome analysis in order to investigate the seed storage proteins. A systematic proteome profiling was conducted through one-dimensional gel electrophoresis followed by MALDI-TOF-TOF mass spectrometry in the seeds (cotyledonary tissue) of soybean genotypes. Two dimensional gels stained with CBB, a total of 10 proteins were identified and analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. A total of ten proteins such as glycinin Gy4 precursor, glycinin G3 precursor, glycinin G1 precursor, glycinin chain A2B1a precursor, glycinin chain A2B1a precursor were identified in our investigation. However, the glycinin subunit may be considered to play important roles in soybean breeding and biochemical characterization. In addition, the improved technique will be useful to dissect the genetic control of glycinin expression in soybean.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.648-657
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• 2008

In this study, we used functional genomic strategies, proteomics and quantitative real-time (qRT)-PCR, to advance our understanding of genes associated with fumonisin production in the fungus Fusarium verticillioides. Earlier studies have demonstrated that deletion of the FCC1 gene, which encodes a C-type cyclin, leads to a drastic reduction in fumonisin production and conidiation in the mutant strain (FT536). The premise of our research was that comparative analysis of F. verticillioides wild-type and FT536 proteomes will reveal putative proteins, and ultimately corresponding genes, that are important for fumonisin biosynthesis. We isolated proteins that were significantly upregulated in either the wild type or FT536 via two-dimensional polyacrylamide gel electrophoresis, and subsequently obtained sequences by mass spectrometry. Homologs of identified proteins, e.g., carboxypeptidase, laccase, and nitrogen metabolite repression protein, are known to have functions involved in fungal secondary metabolism and development. We also identified gene sequences corresponding to the selected proteins and investigated their transcriptional profiles via quantitative real-time (qRT)-PCR in order to identify genes that show concomitant expression patterns during fumonisin biosynthesis. These genes can be selected as targets for functional analysis to further verify their roles in $FB_1$ biosynthesis.