• Title/Summary/Keyword: G- protein

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Fabrication of Protein A-Viologen Hetero LB Film for Antibody Immobilization

  • Lee, Heon-Ju;Choe, Jeong-U;Lee, U-Chang;O, Byeong-Geun;Lee, Won-Hong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.859-862
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    • 2001
  • For the development of preferable immunosensor and protein chip, the viologen Langmuir-Blodgett (LB) multilayer was fabricated on the surface, and then protein A was adsorbed on the proposed viologen LB film by electrostatic attractive force. The Immunoglobulin G (IgG) labeled with fluorescence marker was self-assembled on the fabricated protein A film. The topographies of the deposited films were investigated by using atomic force microscope (AFM). The immobilization of IgG was verified by fluorescence spectrum. Such structures can be used as sublayers for various kinds of IgG immobilization toward immunosensors and protein chip.

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Escherichia coli GroEL was Induced by the Expression of the Cloned Bacillus megaterium ATCC14945 Pencillin G Acylase Gene (클론된 Bacillus megaterium ATCC14945의 페니실린 지 아실라제의 발현에 따른 대장균에서의 GroEL의 유도 생산)

  • Hyun, Kang Joo;Kim, Sung Sun;Yoo, Ook Joon
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.421-424
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    • 1992
  • Escherichia coli JM83 harboring penicilin G acylase gene of Bacillus megaterium ATCC14945 produced a protein in large amount (>20% of the total protein). The protein was identified as GroEL, one of the E. coli heat shock protein, by N-terminal amino acid sequence analysis. It was found that GroEL was induced by the expressed foreign penicilin G acylase at both 27 and $37^{\circ}C$.

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Protein Quality Evaluation of Cooked Hagfish (Eptatretus burgeri) Meats

  • Hwang, Eun-Young;Lee, Jin-Hwa;Ryu, Hong-Soo;Park, Nam-Gyu;Chun, Soon-Sil
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.287-292
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    • 2002
  • The effect of cooking methods on in vivo and in vivo indices of the protein quality of hagfish meat were investigated. In vivo protein digestibilities of cooked meats (81.3~83.5 %) were not significant different (p<0.05) from those of van meat (82.9%), with the exception of steamed (11$0^{\circ}C$, 15 min) meat (86.3 %). Convection oven cooking (22$0^{\circ}C$, IS min) resulted in a higher trypsin indigestible substrate (TIS, 49.2 mg/g solid) compared with that of raw meat (38.9 mg/g solid). free amino acid content of raw meat was decreased after boiling (10$0^{\circ}C$, 10min). Both convection oven and microwave cooking (2,450 MHz, 3 min) decreased available lysine from 4.9g/16g N to 3.8~4.1g/16g N. In vivo apparent protein digestibilites (AD) of hagfish meat were similar fur raw (92.4%) and cooked meats, but were somewhat lower than ANRC (Animal Nutrition Research Council) casein (945%). The PERs (3.7~4.1) and NPRs (3.7~4.9) of cooked meats were significantly higher (p<0.05) than those of raw meat (PER 3.3, NPR 3.6 and ANRC casein (PER 2.5, NPR 2.6), despite their lower in vivo protein digestibilities. These results demonstrate that cooking at optimal conditions resulted in remarkably positive effects on in vivo and in vivo protein qualities of hagfish meats. Therefore, steamed hagfish meat is an excellent source of high quality protein from seafood products.

Extraction of protein from defatted sesame meal using the enzyme from Bacillus sp. CW-1121 (Bacillus sp. CW-1121이 생성하는 단백 분해 효소를 이용한 참깨박 단백질의 용출)

  • Choi, C.;Chun, S.S.;Cho, Y.J.
    • Applied Biological Chemistry
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    • v.36 no.2
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    • pp.121-126
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    • 1993
  • To extract insoluble proteins of sesame meal residue by using microorganism, the sesame meal residue was treated with crude enzyme solution from Bacillus sp. CW-1121. It was found that the solubility reached to maximum at pH 7.5, $45^{\circ}C$. Under optimum condition, the nitrogen solubility with the enzyme solution from Bacillus sp. CW-1121 reached to 60% in 2 hours. Nitrogen solubility of protein from sesame meal showed minimum value at pH 4.5 and significantly increased above pH 6.0. When the protein from sesame meal extracted with Bacillus sp. CW-1121 was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, water soluble protein was showed 4 bands and salt soluble protein was showed 2 bands. The amino acid composition of water soluble protein, salt soluble protein and free amino acid indicated relatively high contents of serine (17.24 mg/g), glutamic acid (10.77 mg/g) and glutamic acid (6.55 mg/g). Specially, the contents of essential amino acids were high.

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The Localization of the Specific Antigenic Protein in the Tissue of Paragonimus westermani Metacercaria (폐흡충 피낭유충 조직에 있어서 특정항원성 단백질의 분포)

  • Kim, Soo-Jin;Roh, Tae-Hoon;Joo, Kyoung-Hwan;Rim, Han-Jong
    • Applied Microscopy
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    • v.27 no.4
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    • pp.403-416
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    • 1997
  • In order to observe the localization of the specific antigenic protein in the tissue of Paragonimus westermani metacercaria, immunogoldlabeling method was applied using IgG of the dog which were infected with Paragonimus westermani metacercaria and IgG of rabbits which were immunized with purified 23 kDa protein from metacercaria of the Paragenimus westermani. The metacercaria worm tissues obtained from Cambaroides similis were embedded in Lowicryl HM20 medium, treated with infected and immunized IgG and protein A gold complex (particle size; 12 nm) and observed by electron microscope. In the tissue antigen of Paragonimus westermani metacercaria, the content of excretory bladder which was highly dense electron density was constituted in the excretory bladder of the parenchymal tissue. In the metacercaria tissues antigen reacted with IgG of infected dog. Labeled gold particles distributed on the interstitial matrix of parenchymal cells, fibrous granules of parenchymal tissue and the content of excretory bladder. High antigenicity was observed on content of excretory bladder. It was found to be specifically distributed at the tissue of Paragonimus westermani metacercaria. In the tissues antigen reacted with IgG of immunized rabbit. Labeled gold particles randomly distributed on the interstitial matrix and fibrous granules of parenchymal tissue but in the content of excretory bladder of Paragonimus westermani metacercaria, gold particles were richly labeled. Therefore, the 23 kDa protein contained with Paragonimus westermani metacercaria was found protein which was specifically constituted at the content of excretory bladder of Paragonimum westermani metacercaria. The 23 kDa protein was commonly contained from of Paragonimus westermani metacercaria to adult and showed strong antigenicity against the immunized and infected IgG.

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Dietary protein requirements of abalone (Haliotis discus, Reeve 1846) depending on abalone size

  • Baek, Seong Il;Cho, Sung Hwoan
    • Fisheries and Aquatic Sciences
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    • v.24 no.3
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    • pp.129-137
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    • 2021
  • Dietary protein requirements of abalone (Haliotis discus) depending on abalone size were determined and compared. One thousand and fifty small abalone (initial weight of 2.7 g) and five hundred forty large one (initial weight of 16.0 g) were distributed into 15 and 18 containers in Trial 1 and 2, respectively. Five and six experimental diets containing crude protein level from 20% to 40% and 20% to 45% with 5% increment of protein level for the small and large abalone were prepared and referred to as the CP20, CP25, CP30, CP35, CP40, and CP45 diets, respectively. The experimental diets were fed to abalone for 16 weeks in Trials 1 and 2. Specific growth rate (SGR) of the small abalone fed the CP20 diet was lower compared to that of abalone fed all other diets in Trial 1. Growth performance (weight gain and SGR) of the large abalone fed the CP30, CP35, and CP40 diets were greater than that of abalone fed the CP20, CP25, and CP45 diets in Trial 2. Dietary protein requirements were estimated to be 33.0% and 33.5% for the small and large abalone based on the 2nd order polynomial analysis, respectively. Dietary protein requirements for the small abalone grown from 2.7 g to 7.4 g and the large one grown from 16 g to 21 g were estimated to be 33.0% and 33.5%, respectively. Size differences in abalone did not affect dietary protein requirement under this experimental conditions.

Nitrogen Metabolism in Lactating Goats Fed with Diets Containing Different Protein Sources

  • Santos, A.B.;Pereira, M.L.A.;Silva, H.G.O.;Pedreira, M.S.;Carvalho, G.G.P.;Ribeiro, L.S.O.;Almeida, P.J.P.;Pereira, T.C.J.;Moreira, J.V.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.5
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    • pp.658-666
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    • 2014
  • This study aimed to evaluate urea excretion, nitrogen balance and microbial protein synthesis in lactating goats fed with diets containing different protein sources in the concentrate (soybean meal, cottonseed meal, aerial part of cassava hay and leucaena hay). Four Alpine goats whose mean body weight was $42.6{\pm}6.1kg$ at the beginning of the experiment, a mean lactation period of $94.0{\pm}9.0days$ and a production of $1.7{\pm}0.4kg$ of milk were distributed in a $4{\times}4$ Latin square with four periods of 15 days. Diets were formulated to be isonitrogenous, containing 103.0 g/kg of CP, 400 g/kg of Tifton 85 hay and 600 g/kg of concentrate. Diet containing cottonseed meal provided (p<0.05) increased excretion of urea and urea nitrogen in the urine (g/d and mg/kg of BW) when compared with leucaena hay. The diets affected the concentrations of urea nitrogen in plasma (p<0.05) and excretion of urea nitrogen in milk, being that soybean meal and cottonseed meal showed (p<0.05) higher than the average aerial part of the cassava hay. The use of diets with cottonseed meal as protein source in the concentrate in feeding of lactating goats provides greater nitrogen excretion in urine and negative nitrogen balance, while the concentrate with leucaena hay as a source of protein, provides greater ruminal microbial protein synthesis.

Isolation of GTP Binding Protein from Bovine Brain (소의 뇌로부터 GTP 결합단백질의 분리)

  • Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.360-368
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    • 1993
  • GTP binding protein (G-protein) associated with membrane and involved in signal transduction was isolated from bovine brain, and molecular weight of G protein was observed. As the results, cell membranes were homogenized from bovine brain tissues and proteins of membrane were gained using 1% cholate, and progressed the chromatography. The purification process was performed by step, DEAE-Sephacel, Ulttrogel AcA 34 and heptylamine-Sepharose column chromatography. The chromatographic fractions were confirmed by GTP binding assay and SDS-polyacrylamide gel electrophoresis. Molecular weight of $Go{\alpha}$ was revealed 39,000 dalton and $G{\beta}$ 36,000 dalton. One more step of heptylamine-Sepharose was enforced to purify the GTP binding protein. Finally I gained the GTP binding protein isolated subtype of $Go{\alpha}$ and $G{\beta}$.

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Structural Features of β2 Adrenergic Receptor: Crystal Structures and Beyond

  • Bang, Injin;Choi, Hee-Jung
    • Molecules and Cells
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    • v.38 no.2
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    • pp.105-111
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    • 2015
  • The beta2-adrenergic receptor (${\beta}2AR$) belongs to the G protein coupled receptor (GPCR) family, which is the largest family of cell surface receptors in humans. Extra attention has been focused on the human GPCRs because they have been studied as important protein targets for pharmaceutical drug development. In fact, approximately 40% of marketed drugs directly work on GPCRs. GPCRs respond to various extracellular stimuli, such as sensory signals, neurotransmitters, chemokines, and hormones, to induce structural changes at the cytoplasmic surface, activating downstream signaling pathways, primarily through interactions with heterotrimeric G proteins or through G-protein independent pathways, such as arrestin. Most GPCRs, except for rhodhopsin, which contains covalently linked 11 cis-retinal, bind to diffusible ligands, having various conformational states between inactive and active structures. The first human GPCR structure was determined using an inverse agonist bound ${\beta}2AR$ in 2007 and since then, more than 20 distinct GPCR structures have been solved. However, most GPCR structures were solved as inactive forms, and an agonist bound fully active structure is still hard to obtain. In a structural point of view, ${\beta}2AR$ is relatively well studied since its fully active structure as a complex with G protein as well as several inactive structures are available. The structural comparison of inactive and active states gives an important clue in understanding the activation mechanism of ${\beta}2AR$. In this review, structural features of inactive and active states of ${\beta}2AR$, the interaction of ${\beta}2AR$ with heterotrimeric G protein, and the comparison with ${\beta}1AR$ will be discussed.

Blood biochemical parameters and organ development of brown layers fed reduced dietary protein levels in two rearing systems

  • Viana, Eduardo de Faria;Mello, Heloisa Helena de Carvalho;Carvalho, Fabyola Barros;Cafe, Marcos Barcellos;Leandro, Nadja Susana Mogyca;Arnhold, Emmanuel;Stringhini, Jose Henrique
    • Animal Bioscience
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    • v.35 no.3
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    • pp.444-452
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    • 2022
  • Objective: An experiment was conducted to evaluate the effect of different levels of crude protein (CP) and two rearing systems (cage and floor), on blood parameters and digestive and reproductive organ development of brown laying hens. Methods: A total of 400 Hisex Brown laying hens between 30 and 45 weeks of age were distributed in a completely randomized design and a 2×4 factorial arrangement, with main effects including two rearing systems (cage and floor) and levels of CP (140, 150, 160, and 180 g/kg), in a total of eight treatments and five replicates of 10 birds each with initial body weight of 1,877 g (laying hen in cage) and 1,866 g (laying hens in floor). The parameters evaluated were plasma total protein, albumin, uric acid, total cholesterol, relative weights of oviduct, abdominal fat, liver, gizzard, crest and dewlap, length of small intestine and oviduct. Results: The blood parameters were similar in birds reared in cage and floor systems. The birds reared on the floor showed greater small intestine and oviduct weight (%) and lower liver and pancreas weight (%). A significant interaction was observed between factors for the relative gizzard, crest and dewlap weight, serum protein, uric acid, and total cholesterol (p<0.05). The diets with 140 g/kg CP resulted in lower serum protein and lower cholesterol in birds reared in floor system, while birds reared in cage system showed no effect of CP on both parameters. Birds reared in cage and fed with 140 and 150 g/kg CP presented lower uric acid. The group of birds reared in floor system fed 180 g/kg had greater uric acid. Conclusion: The dietary protein level can be reduced up to 140 g/kg for Hisex Brown hens (30 to 45 weeks of age) without an important effect on metabolic profile and organ development in both rearing systems.