• KSBB Journal
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• 제30권6호
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• pp.296-301
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• 2015

Eggshell-based biocomposites have become attractive due to their exquisite nanostructure and biological properties, which are mainly composed of highly organized calcium carbonate crystals controlled by organic macromolecules such as proteins and polysaccharides. Here, we designed the recombinant fusion protein of a putative eggshell matrix protein named as GG1234 and a fluorescent reporter protein of DsRed. The protein was successfully over-expressed in E. coli and purified by Ni-NTA affinity chromatography. In vitro calcium carbonate crystallization was conducted in the presence of the fusion protein, and morphological change was investigated. The protein inhibited the calcite growth in vitro, and spherical calcium carbonate micro-particles with the diameter of about $20-30{\mu}m$ were obtained. We expect that this study would be helpful for better understanding of eggshell-based biomineralization.

• Journal of Nutrition and Health
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• 제16권2호
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• pp.115-124
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• 1983

The biochemical nature of the protein constituents of six year old fresh Panax ginseng root was studied. Total protein constituents were extracted with phosphate buffer of pH 7.4, ionic strength of 0.1 and fractionated by ultrafiltration using four different membranes which cut down the materials of molecular weight of 500, 1,000, 5,000 and 10,000, respectively. Each fraction was subjected to ion exchange chromatography using DEAE - cellulose to isolate component proteins. The protein fraction larger than molecular weight of 10,000 was refractionated by the method of ammonium sulfate precipitation. The electrophoresis of the refractionated protein constituents was performed. The amino acid composition of the protein constituents was determined by gas- liquid chromatography. From the results, it could be summarized that eleven different protein constituents smaller than molecular weight of 10,000 were isolated from the fresh Panax ginseng root. At least eleven different protein constituents larger than molecular weight of 10,000 were identified from the electrophoretic patterns. These protein costituents seem to be compounded of all or some of five different subunits.

• 한국자기공명학회논문지
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• 제21권1호
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• pp.13-19
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• 2017

Human sterol ${\Delta}8-{\Delta}7$ isomerase, commonly known as emopamil binding protein (EBP), is an essential protein in the cholesterol-synthetic pathway, and mutations of this protein are critically associated with human diseases such as Conradi-Hunermann-Happle or male EBP disorder with neurological defects syndrome. Due to such a clinical importance, EBP has been intensively investigated and some important features have been reported. EBP is a tetra-spanning membrane protein, of which $2^{nd}$, $3^{rd}$, and $4^{th}$ membrane-spanning ${\alpha}$ helices play an important role in its enzymatic function. However, detailed structural feature at atomic resolution has not yet been elucidated, due to characteristic difficulties in dealing with membrane protein. Here, we over-expressed EBP using Escherichia coli and performed detergent screening to find suitable membrane mimetics for structural studies of the protein by NMR. As results, DPC and LMPG could be evaluated as the most favorable detergents to acquire promising NMR spectra for structural study of EBP.

• BMB Reports
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• 제31권4호
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• pp.350-354
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• 1998

To understand the role of protein kinase C (PKC) in the regulation of chondrogenesis, we examined proteins which are phosphorylated by PKC. Stage 23/24 chick embryo wing mesenchymes were micromass-cultured to induce chondrogenesis and cell extracts were phosphorylated in a condition that activates PKC. Several proteins including 63 and 66 kDa proteins were phosphorylated. The 66 kDa protein was phosphorylated only in the presence of phorbol 12-myristate 13-acetate (PMA) and phosphatidylserine CPS), and the phosphorylation was almost completely diminished by bisindolylmaleimide, a PKC inhibitor. In addition, partially purified PKC increased the phosphorylation of the 66 kDa protein. Treatment of cultures with lysophosphatidylcholine (LPC) promoted chondrogenesis and phosphorylation of 66 kDa protein, while PMA and thymeleatoxin inhibited both of the two events. Our results suggest that the 66 kDa protein is a putative substrate of PKC, and phosphorylation of the 66 kDa protein, probably by $PKC\alpha$ is required for chondrogenesis.

• Journal of Photoscience
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• 제7권3호
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• pp.123-128
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• 2000

Phosphorylation of cellular proteins is a key regulatory mehanism for signal transduction pathway in living cells. Phytochrome, a red/far-red light photoreceptor in plants, is known to employ protein phosphorylation for its light signaling, although its detauked mechanism is still ambiguous. This study is intended to identify the phosphoproteins and protein kinases that are regulated by phytochrome, by employing transgenic rice seedlings that overexpress Arabidopsis phytochrome A. Red light stimulated phsophorylation of a 70 kDa protein and far-red light negated the effect. The red light induced phosphotylation of the 70 kDa protein was strongly activated by heparin and inhibited by poly-L-lysine, suggesting that the 70 kDa protein phosphorylating kinase belongs to GSK-3/SHAGGY protein kinase that has functional roles in establishing cell fate and pattern formation in Drosophila. Taken together with the fact that phytochrome controls plant development, these results may suggest that a GSK-3/SHAGGY-related protein kinase in plant(ASK) is likely to be involved in phytochrome signal transduction.

• Journal of Microbiology and Biotechnology
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• 제16권7호
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• pp.1026-1031
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• 2006

The optimization of the production of a fusion protein containing the antigenic domain 1 (AD-1) is of a great importance, considering its use in diagnostic tests. The fusion protein is produced by the fermentation of a recombinant strain of Escherichia coli containing the plasmid Mbg58, which expresses the AD-1 (aa 484-650) of human cytomegalovirus glycoprotein B as a fusion protein together with aa 1-375 of ${\beta}-galactosidase$. An important characteristic of promoters (lac and derivatives) used in recombinant protein production in E. coli is their inducibility. Induction by IPTG is widely used for basic research; however, its use in large-scale production is undesirable because of its high cost and toxicity. In this work, studies using different inducers and carbon sources for the production of a fusion protein containing the AD-l were performed. The results showed that lactose could be used as an inducer in the fermentation process for the production of this protein, and that expression levels could exceed those achieved with IPTG. The use of lactose for protein expression in E. coli should be extremely useful for the inexpensive, large-scale production of heterologous proteins in E. coli. Addition of sucrose to the fermentation medium improved the yield of recombinant protein, whereas addition of fructose or trehalose decreased the yield.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1022-1027
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• 2005

Based on plasmid display technology by the complexes of fusion protein and the encoding plasmid DNA, an in vitro selection method for high affinity DNA-binding protein was developed and experimentally demonstrated. The GAL4 DNA-binding domain (GAL4 DBD) was selected as a model DNA-binding protein, and enhanced green fluorescent protein (EGFP) was used as an expression reporter for the selection of target proteins. Error prone PCR was conducted to construct a mutant library of the model. Based on the affinity decrease with increased salt concentration, mutants of GAL4 DBD having high affinity were selected from the mutant protein library of protein-encoding plasmid complex by this method. Two mutants of (Lys33Glu, Arg123Lys, Ile127Lys) and (Ser47Pro, Ser85Pro) having high affinity were obtained from the first generation mutants. This method can be used for rapid in vitro selection of high affinity DNA-binding proteins, and has high potential for the screening of high affinity DNA-binding proteins in a sequence-specific manner.

• Journal of Nutrition and Health
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• 제29권8호
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• pp.908-915
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• 1996

Protein concentration in human from 39 well-norished American women and its adequacy for growth of exclusively breastfed infants(BF) and breastfed infants fed supplementary foods(BFS)from 1-6 months postpartum were studied. Mean protein concentration of breast milk measured by Lowry et al., using human serum albumin as a standard, over the first 6 months lactation was 1.31$\pm$0.13g/dl. Concentration of protein was singnificantly higher at the first month of lactation (1.55$\pm$0.23g/dl)(P<0.05) than any other month studied. Mean volume of breast milk ranged from 662-848ml/day in the BE group and from 415-661ml/day in the BFS group during the first 6 months of lactation. Mean protein intake of infants ranged from 1.3-2.2g/kg in the BF group and from 1.4-2.1g/kg in the BFS group. Mean protein intake (g/kg body weight) of both BF and BFS groups was less than Recmmended Dietary Allowance(1989, USA) of 2.2g/kg except at 1 month of age. However, mean growth of the infants was normal according to NCHS reference, suggesting that the RDA for protein was unrealistically high for infants during 2-6 months of age. Protein provided by breast milk alone appeared adequate for normal growth during this time.

• 정보처리학회논문지A
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• 제16A권3호
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• pp.153-158
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• 2009

구조적 생물 정보학 분야는 단백질의 3차원 구조를 대상으로 단백질을 연구하는 분야이며, 구조적생물 정보학의 중요한 분야 중의 하나는 단백질 3차원 구조 가시화이다. 단백질의 3차원 구조를 규명하는 장비의 발달로, 규명되는 단백질의 크기와 개수가 증가함에 따라, 고성능의 단백질 가시화 시스템의 필요성도 크게 증가하였으나, 기존의 단백질 구조 가시화 시스템은 3차원 그래픽 하드웨어에 최적화 되지 못하여, 거대 단백질의 가시화에 충분한 성능을 가지지 못하였다. 본 논문에서 제안하는 단백질 3차원 구조 가시화 시스템은 거대 단백질의 가시화 하기 위하여, 3차원 그래픽 하드웨어의 최적화 기법중의 하나인 기하 인스턴싱 기법을 사용하여 빠르게 거대 단백질을 렌더링 한다. 성능 실험에서 7종의 다른 크기의 단백질을 대상으로, 4가지 가시화 방법에 대하여, 제안하는 시스템과 기존의 시스템과의 단백질 렌더링 성능 비교 실험을 하여, 대부분의 경우 우수한 성능을 보였다.

• 대한수의학회지
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• 제6권1호
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• pp.37-41
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• 1966

Observations were made on the total serum protein values of 532 Korean cattles(341 females and 191 males and 191 males) and 90 milk cows in order to determine the effects of age, sex and nutritional status upon serum protein. Results obtained in this studies were summerized as follows; 1. The concentration of total serum protein was higher in the adult than in the younger and was generally increased with the advance of age in Korean cattle. The rate of increase in total serum protein content was more significant in the young than that of adult. 2. The average concentration of total serum protein of male did not differ significaltly from that of females in Korean cattle. 3. The concentration of total serum protein in milk cows were significaltly higher than that of Korean cattle cows and this difference seemed to be influenced by rations high in protein 4. A general tendency showed that the concentration of total serum protein was significaltly higher in the Korean cattle keeping with good nutritional status than that of poor nutritional status.