• 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.

• 약학회지
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• 제54권3호
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• pp.180-186
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• 2010

Aldosterone, mineralocorticoid hormone, has important functions related to the regulation of blood pressure and balance of fluids and electrolytes in the distal region of the nephron. By genomic and non-genomic action of aldosterone, the physiological kidney functions are modulated. However, many of them except several kind of sodium channel have not been identified and analyzed yet. In this study, proteomic technologies with two-dimensional gel electrophoresis (2-DE) gel using aldosterone rat model were applied to analyze and identify the aldosterone dependently expressed proteins in rat kidney cortex. As a result, the established aldosterone rat model exhibited the normal physiological responses to aldosterone and modulated proteins were identified, which included 15 increased and 3 decreased proteins on 2-DE analysis. Among them, 11 proteins were identified as changed proteins by LC-MS/MS analysis. These proteins identified as aldosterone induced proteins were involved in several cellular pathways such as cytoskeleton remodeling, energy metabolism, amino acid metabolism, and chaperone process. In conclusion, our data could provide the insights into the new mechanism underlying regulation of kidney functions by aldosterone.

• 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.

• 한국축산식품학회지
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• 제38권5호
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• pp.1101-1108
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• 2018

The aim of this study was to investigate the effects of dry-aging (DA) and the cooking process on the myofibril protein functionalities and in vitro digestibility of proteins in beef loin. Six sirloins from beef were dry-aged for 28 d, and the control group (n=6) was analyzed 2 d postmortem for this study. Dimensional changes (reduction of thickness and surface shrinkage) after cooking were significantly greater in the control group than the DA group, whereas the shear force of the DA group was significantly lower than that of the control. Effect of cooking on aggregation, hydrophobicity, and in vitro digestibility were significantly higher in the DA group than in the control. After cooking, the protein in DA sirloins was more oxidized than in the control samples. According to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis result, the low molecular weight bands (below 17 kDa) increased in the DA group, finding that the protein characteristics of dry-aged beef was affected by cooking.

• 한국동물학회지
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• 제28권2호
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• pp.71-84
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• 1985

성게의 일종인 Hemicentrotus pulcherrimus에서 수정전에 이미 축적되었던 mRNA의 활성화 변화를 알아보기 위하여 초기 발생동안의 RNA와 단백질 합성에 관하여 연구하였다. 미수정란에서는 RNA와 단백질의 합성들이 대단히 낮았다. 그러나 수정과 함께 RNA합성은 크게 변하지 않은 반면, 단백질합성은 크게 활성화되었다. RNA와 단백질 합성율이 병행적으로 변하지는 않지만, 포배와 낭배에서 크게 증가함을 볼 수 있었다. 단백질합성은 양적으로 뿐만 아니라 질적으로도 발생단계에 따라 변하는 사실을 이차원 전기영동에 의한 연구를 통하여 확인할 수 있었다.

• Food Science and Biotechnology
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• 제14권6호
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• pp.854-859
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• 2005

Although action modes of equol and genistein have been extensively studied, their precise roles in tumor cells remain elusive. To address possible effects of these compounds on protein expression in mammary tumor cells, proteins modulated in MCF-7 mammary tumor cells when incubated in absence and presence of 10 uM equol or genistein were identified through 2-dimensional gel electrophoresis, MALDI-TOF MS/MS, and NCBInr database search using Mascot software. Most proteins differentially expressed in MCF-7 cells after treatment with 10 uM genistein or equol were identified as being the same. Exposure to both compounds caused decreased cellular expression of RNA-binding protein regulatory subunit and oncogene DJ1 tubulin beta-1 chain, and increased expression of heterogeneous ribonucleoproteins F and L, KH-type splicing regulatory protein, and translation elongation factor EF-Tu precursor. Genistein and equol at dose used in this study showed common action mechanism.

• Reproductive and Developmental Biology
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• 제36권2호
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• pp.121-131
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• 2012

After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q-ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.

• 한국양식학회지
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• 제10권3호
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• pp.301-310
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• 1997

황해서식 두족류 (class Cephalopoda)의 가용성 단백질에 대한 연구를 위해, 인천 및 목포 연근해에서 채집된 두족류 3목 5종의 (오징어목 : 참갑오징어 (Sepia esculenta) 및 쇠갑오징어 (Sepiella japonica), 살오징어목 : 한치꼴뚜기 (Loligo chinensis) 및 참꼴뚜기 (Loligo beka), 문어목 : 낙지 (Octopus minor)의 안구단백질, 근육단백질 및 간조직을 추출하여, 각종 전기영동 (Davis-PAGE 및 SDS-PAGE, Exponential gradient SDS-PAGE, 등전점 전기영동, 2차원 전기영동) 방법에 의한 단백질 분리 양상을 통해 두족류 종사이의 유전적 근연관계를 분석하였다. 시료의 안구 및 근육 단백질에 대한 exponential gradient SDS-PAGE 전기영동 결과 대략 분자량 35-50 KDa 사이에서 단백질 분리 양상의 차이를 볼 수 있었으며, 등전점 전기영동 방법(IEF)에 의해서는 pI값 7.5-8.5 사이에서 종간 특이성을 갖는 단백질 분리 양상을 볼 수 있었다. 특히 유의성이 있다고 판단된 시료의 안구 단백질을 2차원 전기영동 방법에 의해 분리 해 본 결과 대부분 분자량 30-50 KDa 사이에 분포하고 있어 exponential gradient SDS-PAGE 전기 영동에 의한 결과와 일치함을 알 수 있었다.

• 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.

• 한국가축번식학회지
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• 제16권4호
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• pp.289-299
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• 1993

돼지 난포내의 각 구성분들에 대해 10% SDS-PAGE와 IEF를 이용한 이차원 전기영동을 실시하여 세포 및 난포액 구성분의 구조단백질상을 분석하였다. 난자-난구세포 복합체를 호르몬과 15%의 FCS가 포함된 M16 배양액으로 39$^{\circ}C$, 5% CO2 상태에서 35시간 동안 체외배양하였다. 배양 전후의 난자, 투명대 및 난구세포와 난포 크기별로 회수된 난포액들을 각각 분리 회수하여 구조단백질상을 분석하였으며, Silver 염색과 CBB 염색으로 분석이 가능한 각 구성분의 적정 시료량을 조사하였다. 한편 난포 구성분들에 있어서 난자는 분자량이 25와 114kd, 난구세포는 20, 33, 58, 78 및 112kd, 투명대는 65kd, 그리고 난포액은 18, 76, 92, 152 및 187kd 단백질을 세포특이단백질로 가지고 있음이 확인되었다. 특히 난자의 경우 성숙에 따라 구조단백질상의 변화가 확인된 반면, 난구세포에서는 차이가 없었다. 또한 난포액은 난포의 크기에 따라서는 단백질상의 차이가 없었으나 호르몬 처리 여부에 따라서는 이차원 전기영동상에서 몇가지 단백질에서 차이가 확인되었고, 난포세포들도 폐쇄 여부에 따라 단백질 조성에 차이를 보였다. 따라서 본 실험에서는 전기영동에 필요한 시료의 양과 준비 방법을 확립하여 각 난포 구성분들의 단백질상 분석에 대한 기초자료를 확립하였으며, 이상의 결과는 앞으로 진행될 단백질의 생합성 분석이나 면역화학학적 분석에 유용하게 이용될 수 있을 것이다.