• 센서학회지
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• 제20권6호
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• pp.434-438
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• 2011

A portable surface plasmon resonance(SPR) based immunosensor using thiolated protein G and protein G was developed for the detection of immunoglobulin G(IgG). The protein G has specific affinity with Fc fragment of IgG and was thiolated by 2-Iminothiolane for introduction of thiol groups. Anti-IgG, bovine serum albumin(BSA), and IgG have been sequently injected after surface modification of gold sensor chip with protein G and thiolated protein G. The output signal was increased with the injection of each protein and the actual signal was measured by subtracting signal of reference channel from signal of sample injected channel. The experimental results showed the higher detection capability of IgG using thiolated protein G compared with protein G. From these results, we can conclude that the current surface modification technique and the portable SPR sensor system can be applied to various immunosensors for diagnosis.

• Toxicological Research
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• 제29권3호
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• pp.149-155
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• 2013

G protein-coupled receptors (GPCRs) are membrane receptors; approximately 40% of drugs on the market target GPCRs. A precise understanding of the activation mechanism of GPCRs would facilitate the development of more effective and less toxic drugs. Heterotrimeric G proteins are important molecular switches in GPCR-mediated signal transduction. An agonist-activated receptor interacts with specific sites on G proteins and promotes the release of GDP from the $G{\alpha}$ subunit. Because of the important biological role of the GPCR-G protein coupling, conformational changes in the G protein upon receptor coupling have been of great interest. One of the most important questions was the interface between the GPCR and G proteins and the structural mechanism of GPCR-induced G protein activation. A number of biochemical and biophysical studies have been performed since the late 80s to address these questions; there was a significant breakthrough in 2011 when the crystal structure of a GPCR-G protein complex was solved. This review discusses the structural aspects of GPCR-G protein coupling by comparing the results of previous biochemical and biophysical studies to the GPCR-G protein crystal structure.

• 센서학회지
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• 제19권2호
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• pp.142-148
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• 2010

We have compared and investigated the detection capabilities of antibody of immunoglobulin G(anti-IgG) immobilized by protein G and N-hydroxysuccinimide(NHS) at the end of the self-assembled monolayer(SAM). Surface plasmon resonance(SPR) sensor has been utilized to measure the interaction between biomolecules. After formation of the protein G and SAM, anti-IgG, bovine serum albumin(BSA) and IgG has been sequently injected. Through the reponse of the SPR, we can conclude that the protein G immobilized anti-IgG better than the SAM. In addition, IgG detection capability of the anti-IgG immobilized by the protein G showed better performance compared with that immobilized by the SAM.

• BMB Reports
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• 제33권2호
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• pp.188-192
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• 2000

Recombinant protein G has been utilized in the purification of antibodies from various mammalian species based on the interaction of antibodies with protein G. The interaction between immunoglobulin and protein G may not be restricted to the Fc protion of antibodies, as many different $F(ab)_2$ or Fab fragments can also bind to protein G. I found both FAb $F(ab)_2$ of 145-2C11, a hamster anti-mouse $CD3{\varepsilon}$ antibody, bound to the protein G-sepharose. Interestingly, Fab and $F(ab)_2$ of 145-2C11 did not bind to the protein A-sepharose. The binding of Fab and $F(ab)_2$ of 145-2C11 to protein G provided a useful method to remove proteases, chopped fragments of the Fc region, and other contaminating proteins. The remaining intact antibody in the protease reaction mixture can be removed by using a protein A-sepharose, because the Fab and $F(ab)_2$ portions of 145-2C11 did not bind to protein A-sepharose. The specific binding of Fab and $F(ab)_2$ portions of 145-sC11 to a protein G-sepharose (though not to a protein A-sepharose) and binding of intact 145-2C11 to both protein A- and G-sepharose will be useful in developing an effective purification protocol for Fab and $F(ab)_2$ portions of 145-2C11.

• Journal of Microbiology
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• 제44권2호
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• pp.145-154
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• 2006

Heterotrimeric G proteins (G proteins) are conserved in all eukaryotes and are crucial components sensing and relaying external cues into the cells to elicit appropriate physiological and biochemical responses. Basic units of the heterotrimeric G protein signaling system include a G protein-coupled receptor (GPCR), a G protein composed of ${\alpha},\;{\beta},\;and\;{\gamma}$ subunits, and variety of effectors. Sequential sensitization and activation of these G protein elements translates external signals into gene expression changes, resulting in appropriate cellular behaviors. Regulators of G protein signaling (RGSs) constitute a crucial element of appropriate control of the intensity and duration of G protein signaling. For the past decade, G protein signaling and its regulation have been intensively studied in a number of model and/or pathogenic fungi and outcomes of the studies provided better understanding on the upstream regulation of vegetative growth, mating, development, virulence/pathogenicity establishment, and biosynthesis of secondary metabolites in fungi. This review focuses on the characteristics of the basic upstream G protein components and RGS proteins, and their roles controlling various aspects of biological processes in the model filamentous ascomycete fungus Aspergillus nidulans. In particular, their functions in controlling hyphal proliferation, asexual spore formation, sexual fruiting, and the mycotoxin sterigmatocystin production are discussed.

• 한국가금학회지
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• 제30권2호
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• pp.101-106
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• 2003

육용종계 산란기의 적정 단백질공급수준을 구명하기 위하여 종계형 케이지에 아바에이카 육용종계 400 수를 공시하여 40 주간(24∼64주령) 1일 1수당 단백질 급여량을 18, 20 22, 24g 수준으로 사양시험을 실행하였으며 그 결과는 다음과 같다. 1) 총 산란율과 종란 산란율은 모두 처리간에 유의적인 차이가 있었으며(P<0.05), 20g 공급구가 가장 높았으나 18∼22g 공급구간에는 유의차가 없었고 24g 공급구는 20g 공급구에 비해 유의적으로 낮았다{P<0.05). 2) 평균난중은 l 일 단백질공급수준이 증가할수록 증가하는 경향이었으나{P<0.05), 20∼24g 공급구간에는 유의차가 없었고 18g 공급구만 24g 공급구와 유의적인 차이가 있었다. 성계생존율은 처리간에 유의적인 차이가 없었다. 3) 계란 kg당 또는 계란 l개당 사료요구율과 ME요구량은 모두 l일 CP 20g 공급구가 가장 우수했으며(P<0.05), 계란 kg당 사료요구율과 ME 요구량에서는 CP 18g 공 급구 계란1개당 사료요구율과 ME요구량에서는 CP 24g 공급구만 CP 20g 공급구와 유의차를 보였다(P＜0.05). 4) 계란 kg당 또는 계란 l개당 CP 요구량은 모두 l일 단백질공급량이 많아질수록 증가하였으내(P<0.05), 18g과 20g공급구간에는 유의차가 없었다. 5) 계란 kg당 또는 계란 1개당 사료비는 모두 CP 20g공급구가 가장 우수하였으나(P<0.05), 계란 l개당 사료비는 18∼20g 공급구간에 유의차가 없었다. 이상의 결과로 육용종계 산란기의 산란율과 난중, 사료와 영양소이용효율 및 사료비용에서 1일 단백질공급량은 20g이 적정수준이었다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권1호
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• pp.42-47
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• 2005

The possibility of using Moringa oleifera as a ruminant protein supplement was investigated by comparison between nutritive and anti-nutritive value of its different morphological parts with that of conventionally used Leucaena leucocephala leaf meal (LL). Parameters determined were chemical composition, rumen degradable protein (RDP), acid detergent insoluble protein (ADIP), pepsin soluble protein (PESP), non-protein nitrogen (NPN) total soluble protein (TSP) and protein potentially digested in the intestine (PDI). Total phenols (TP) and total extractable tannins (TET) were also evaluated as anti-nutritive factors. In vitro gas production characteristics were measured and organic matter digestibility (OMD) was estimated basing on 24 h-gas production. Crude protein content ranged from 265-308 g/kg DM in M. oleifera leaves (MOL) and seed cake (MOC) respectively. Leucaena leucocephala and Moringa oleifera soft twigs and leaves (MOLSTL) had CP content of 236 and 195 g/kg DM while Moringa oleifera soft twigs alone (MOST) and Moringa oleifera bucks (MOB) had 160, 114 and 69.3 g/kg DM respectively. RDP was highest in (MOC) (181 g/kg DM) followed by (MOL) (177 g/kg DM) and was lowest in MOB (40 g/kg DM). The proportion of the protein that was not available to the animal (ADIP) was (p<0.05) higher in MOL and MOC (72 and 73 g/kg DM) respectively and lowest in LL (29 g/kg DM). The PDI was high in LL (74 g/kg DM) followed by MOC (55 g/kg DM) then MOL (16 g/kg DM). PESP was highest (p<0.05) in MOC followed by MOL then LL (273, 200 and 163 g/kg DM respectively). MOC exhibited highest NPN content (116 g/kg DM) and was lowest in MOB (18 g/kg DM) (p<0.05). Highly (p<0.05) TSP was observed in MOC and MOL (308 and 265 g/kg DM respectively) followed by LL (236 g/kg DM). MOL had negligible TET (20 g/kg DM) when compared with about 70 g/kg DM in LL. Highly (p<0.05) b and a+b values were observed for MOLSTL (602 and 691 g/kg DM respectively) followed by MOL (490 and 538 g/kg DM). Highest c value was observed in MOSTL followed by MOC and MOL (0.064, 0.056 and 0.053 rate/hour) respectively. OMD was highest (p<0.05) for MOSTL followed by MOC and then MOL (579, 579 and 562 g/kg DM respectively). LL exhibited lower (p<0.05) OMD (467 g/kg DM). It was concluded from this study that the high crude protein content in MOL and MOLST could be well utilized by ruminant animals and increase animal performance however, high proportion of unavailable protein to the lower gut of animals and high rumen degradable protein due to negligible tannin content render it a relatively poor protein supplement for ruminants. MOC can be a best alternative protein supplement to leaves and leaves and soft twigs for ruminants.

• Asian-Australasian Journal of Animal Sciences
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• 제23권3호
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• pp.333-339
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• 2010

The objectives of this study were to use the NRC-2001 model and DVE/OEB system to model potential nutrient supply to ruminants and to compare frost damaged (also called "frozen" wheat with normal wheat. Quantitative predictions were made in terms of: i) Truly absorbed rumen synthesized microbial protein in the small intestine; ii) Truly absorbed rumen undegraded feed protein in the small intestine; iii) Endogenous protein in the digestive tract; iv). Total truly absorbed protein in the small intestine; and v). Protein degraded balance. The overall yield losses of the frozen wheat were 24%. Results showed that using the DVE/OEB system to predict the potential nutrient supply, the frozen wheat had similar truly absorbed rumen synthesized microbial protein (65 vs. 66 g/kg DM; p>0.05), tended to have lower truly absorbed rumen undegraded feed protein (39 vs. 53 g/kg DM; p<0.10) and had higher endogenous protein (14 vs. 9 g/kg DM; p<0.05). Total truly absorbed protein in the small intestine was significantly lower (89 vs. 110 g/kg DM, p<0.05) in the frozen wheat. The protein degraded balance was similar and both were negative (-2 vs. -1 g/kg DM). Using the NRC-2001 model to predict the potential nutrient supply, the frozen wheat also had similar truly absorbed rumen synthesized microbial protein (average 56 g/kg DM; p>0.05), tended to have lower truly absorbed rumen undegraded feed protein (35 vs. 48, g/kg DM; p<0.10) and had similar endogenous protein (average 4 g/kg DM; p>0.05). Total truly absorbed protein in the small intestine was significantly lower (95 vs. 108 g/kg DM, p<0.05) in the frozen wheat. The protein degraded balance was not significantly different and both were negative (-16 vs. -19 g/kg DM). In conclusion, both models predict lower protein value and negative protein degraded balance in the frozen wheat. The frost damage to the wheat reduced nutrient content and availability and thus reduced nutrient supply to ruminants by around 12 to 19%.

• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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• pp.718-724
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• 1998

Twelve gilts were used to investigate the effect of lysine to protein ratio (5.2 g lysine/100 g CP vs. 6.7 g lysine/100 g CP) in practical diets on nitrogen retention and the efficiency of utilization in growing pigs. Treatments involved 2 levels of dietary lysine (5.2 or 6.7 g/100 g CP) and 3 levels of dietary crude protein (11, 14 and 17% in diet). Nitrogen retention was greatest when pigs were fed the control diet containing 17% protein. Nitrogen retention progressively increased as dietary protein increased (p < 0.01), but it was not affected by lysine concentration (g/100 g CP). Apparent biological value (ABV, nitrogen retained/apparently digestible nitrogen) was estimated to be ~50% at the maximum nitrogen retention. ABV was not affected by lysine concentration, but declined (p < 0.05) as the dietary protein level increased. The efficiency of intake N used for maximum nitrogen retention was approximately 44%. One gram of lysine supported approximately 9 to 10 g apparent protein accretion (nitrogen retention ${\times}$ 6.25/lysine intake) in pigs fed control diets. The efficiency of lysine utilization for protein accretion was lower in pigs fed high-lysine diets (6.7 g lysine/l00 g CP) so that 1 g of lysine accounted for 7 to 8 g of protein accretion in these pigs (p < 0.01). The lysine required to support maximum nitrogen retention in pigs fed high-lysine diets was higher than that in pigs fed control diets, which suggests that lysine was over-fortified relative to crude protein, since practical diets can not be formulated without excess of some amino acids. In summary the concentration of 5.2 g total lysine/100 g CP in diet is more appropriate for corn-soybean diets than the commonly suggested the content of 6.7 g total lysine/100 g CP.

• 약학회지
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• 제32권5호
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• pp.313-318
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• 1988

Radioprotective ginseng protein fraction was isolated from Korean white ginseng and its cytotoxic effect on CHO-K1 cells was studied by the method of measuring the relative cell survival and total cellular protein content (FRAME method). When ginseng protein at the dose of 300, 600, 900, $1200{\mu}g/ml$ was treated to cells for 24 hrs, the relative survival was significantly decreased at the concentration of above $600{\mu}g/ml$, indicating the presence of cytotoxic effect of the protein at certain concentration. When cellular protein content was measured after ginseng protein at the dose of 300, 600, 900, $1200\;{\mu}g/ml$ was treated, the amount of cellular protein was significantly reduced at the concentration above $600{\mu}g/ml$ in the case of 24 hr treatment and at all concentrations including $300{\mu}g/ml$ in the case of 72 hr treatment. The data suggest that the protein may inhibit cell growth, resulting in the reduction of live cells in culture. $ID_{50}$ value which is the concentration of ginseng protein that reduces the total cellular protein content to 50% of the control was calculated as 2276.86 and $1323.32\;{\mu}g/ml$ in groups treated for 24 and 72 hr, respectively. Since $ID_{50}$ value of above $1000{\mu}g/ml$ indicates very weak cytotoxicity, the ginseng protein seems to exert very weak cytotoxic effect on CHO-K1 cells.