• Title/Summary/Keyword: Protein A

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Human selenium binding protein-1 (hSP56) is a negative regulator of HIF-1α and suppresses the malignant characteristics of prostate cancer cells

  • Jeong, Jee-Yeong;Zhou, Jin-Rong;Gao, Chong;Feldman, Laurie;Sytkowski, Arthur J.
    • BMB Reports
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    • v.47 no.7
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    • pp.411-416
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    • 2014
  • In the present study, we demonstrate that ectopic expression of 56-kDa human selenium binding protein-1 (hSP56) in PC-3 cells that do not normally express hSP56 results in a marked inhibition of cell growth in vitro and in vivo. Down-regulation of hSP56 in LNCaP cells that normally express hSP56 results in enhanced anchorage-independent growth. PC-3 cells expressing hSP56 exhibit a significant reduction of hypoxia inducible protein (HIF)-$1{\alpha}$ protein levels under hypoxic conditions without altering HIF-$1{\alpha}$ mRNA (HIF1A) levels. Taken together, our findings strongly suggest that hSP56 plays a critical role in prostate cells by mechanisms including negative regulation of HIF-$1{\alpha}$, thus identifying hSP56 as a candidate anti-oncogene product.

Hypothetical protein predicted to be tumor suppressor: a protein functional analysis

  • Kader, Md. Abdul;Ahammed, Akash;Khan, Md. Sharif;Ashik, Sheikh Abdullah Al;Islam, Md. Shariful;Hossain, Mohammad Uzzal
    • Genomics & Informatics
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    • v.20 no.1
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    • pp.6.1-6.15
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    • 2022
  • Litorilituus sediminis is a Gram-negative, aerobic, novel bacterium under the family of Colwelliaceae, has a stunning hypothetical protein containing domain called von Hippel-Lindau that has significant tumor suppressor activity. Therefore, this study was designed to elucidate the structure and function of the biologically important hypothetical protein EMK97_00595 (QBG34344.1) using several bioinformatics tools. The functional annotation exposed that the hypothetical protein is an extracellular secretory soluble signal peptide and contains the von Hippel-Lindau (VHL; VHL beta) domain that has a significant role in tumor suppression. This domain is conserved throughout evolution, as its homologs are available in various types of the organism like mammals, insects, and nematode. The gene product of VHL has a critical regulatory activity in the ubiquitous oxygen-sensing pathway. This domain has a significant role in inhibiting cell proliferation, angiogenesis progression, kidney cancer, breast cancer, and colon cancer. At last, the current study depicts that the annotated hypothetical protein is linked with tumor suppressor activity which might be of great interest to future research in the higher organism.

Amino Acid Composition and Nutritional Value of Silkworm Larvae Protein (번데기 단백질(蛋白質)의 아미노산조성과 영양가(營養價)에 관한 연구)

  • Park, Jyung Rewng;Lee, Kyung Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.4
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    • pp.368-373
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    • 1983
  • This study was designed to determine amino acid composition of silkworm larvae protein and to evaluate its nutritional value by rat experiment. Diets were divided into 5 groups; casein ($D_1$), soybean protein ($D_2$), soybean protein+20% silkworm larvae ($D_3$), soybean protein+40% silkworm larvae protein ($D_4$), and silkworm larvae protein ($D_5$). The growth rate, protein efficiency ratio, weight of organs, hematology, and the content of total serum protein and albumin were studied. The amino acid composition of silkworm larvae protein was comparable to the FAO provisional scoring pattern. The protein was especially high in the amount of lysine and methionine indicating that it could be a good supplemental effect for cereals and beans. Growth rate and protein efficiency ratio of silkworm larvae protein were better than soybean protein and these were increased by the addition of silkworm larvae protein. The weight of liver and spleen from silkworm larvae protein group were also higher than soybean protein group and RBC, WBC, Hct and Hb content of 5 groups tested were within the normal ranges. The contents of total serum protein and albumin from soybean protein group were increased by addition of silkworm larvae protein. From the results obtained, it could be stated that the quality of soybean protein might be improved by silkworm larvae protein.

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Selection Attributes and Purchasing Perceptions and Attitudes of Protein Snacks According to Individual Health Lifestyle (개인의 건강 라이프스타일에 따른 단백질 스낵의 선택속성과 구매인식 및 태도)

  • Hwang, Ji Eon;Oh, Ji Eun;Cho, Mi Sook
    • Journal of the Korean Society of Food Culture
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    • v.33 no.4
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    • pp.394-401
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    • 2018
  • This study was conducted to investigate the effects of health lifestyle on high protein snack selection attributes and purchase behaviors among individuals aged 20-30 with high protein snack intake. In addition, the relationship between perception, attitude, satisfaction and recommendation of high protein snacks was invested. Finally, this study aims to provide basic information for marketing high-protein snacks and customized high protein snacks. Analysis of the selection attributes most important for healthy lifestyle, revealed significant differences among all groups excluding the external seeking group (p<0.001). The free living group regarded trust as one of the most important attributes of high protein snack selection, and both the tempered control group and the low-interest group found sensation and price factor to be important. Therefore, when developing high-protein snacks, it is important to determine which attributes of the snack will be highlighted by segmenting the consumer into health lifestyles. Focusing on what ingredients are used to develop high-protein snacks and nutritional ingredients is also important when targeting a free lifestyle group as the main customer. In addition, developing snacks that do not offer depending on the protein content is important when targeting a temperate management group or a low-interest group.

Effect of N-3 Fatty Acids and Dietary Protein Levels on Renal Function in Rats of Different Ages (N-3계 지방산과 단백질 수준이 나이가 다른 흰쥐에서 신장 기능에 미치는 영향)

  • 김화영;정명지;정현주
    • Journal of Nutrition and Health
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    • v.34 no.8
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    • pp.843-849
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    • 2001
  • This study was performed to investigate the effect of n-3 fatty acids and dietary protein levels on renal function. Fifteen-month old male Sprague-Dawley rats were divided into 4 diet groups. Two-month old rats were used as a control group. The experimental diets contained either a% or 25% casein and lipid levels of the diets were 20% by weight. For the control group, the lipid was composed of beef tallow and corn oil on a 1:1 basis, and fish oil was comprised 75% of the fat mixture for the fish oil group. Rats were fed the diets ad libitum for 8 weeks. GFR and urinary protein excretion were higher in high protein groups, while fish oil exhibited no effects. Renal medulla TXB$_2$and PGE$_2$ concentrations tended to be higher in high protein groups and lower in fish oil groups. Light microscopic examinations showed that glomerulosclerosis, tubular atrophy, tubular cast, interstitial inflammation and interstitial fibrosis fended to be higher in aged rats and in high protein groups and lower in fish oil groups. Serum levels of total lipid, triglyceride and total cholesterol were higher in aged rats and lower in fish oil groups while serum HDL-cholesterol level was higher in young rats and in fish oil groups. However, dietary protein level had no effect on serum lipid levels. Serum TBARS concentration was higher in aged rats and in fish oil groups. In conclusion, fish oil caused changes in serum lipid concentrations and eicosanoids metabolism. The effect of fish oil on renal function was less obvious than dietary protein. However, fish oil seemed to be effective in lessening deterioration of renal function due to aging and/or high protein diets through changes in lipid and eicosanoids metabolism.

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Expression and Characterization of Helicobacter pylori Adhesin Protein Linked to Cholera Toxin A2/B Subunits in Escherichia coli

  • Kim, Byung-Oh;Shin, Sung-Seup;Yoo, Young-Hyo;Pyo, Shuk-Neung
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.56-62
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    • 2000
  • The hpa gene genetically linked to the ctxa2b gene was cloned into the pTED expression vector, and the constructed pTEDhpa/ctxa2b was transformed into Excherichia coli. The fusion protein, the adhesin fused to the cholera toxin subunit A2B (CTXA2B) subunit, was expressed to high levels as inclusion bodies in E. coli. The expressed protein was partially purified by washing the inclusion bodies with working solution containing 8M Urea and 0.1M DTT. Refolding of denatured fusion protein was carried out in the presence of glutathione redox buffer. The refolded fusion protein was purified by size exclusion chromatography. The expressed fusion protein was verified by SDS-PAGE, western blotting with antibodies to both antigenic components of adhesin and cholera toxin subunit B (CTXB), and its N-terminal amino acid sequence was analyzed. The orderly assembled fusion protein was confirmed by modified Gm1-ganglioside ELISA with Abs to adhesin. The results indicate that the purified fusion protein is an Adhesin/CTXA2B protein containing the H. pylori adhesin and $G_{m1}4-ganglioside binding activity of CTXB and the expressed fusion protein in E. coli could be easily purified by the refolding process, Its molecular weight was 168kDa as estimated by size exclusion chromatography. The Adhesin/CTXA2B protein may be used as a candidate antigen for oral immunization against H. pylori.

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Heat Shock Protein $90{\beta}$ Inhibits Phospholipase $C{\gamma}-1$ Activity in vitro

  • Cho, Sang-Min;Kim, Sung-Kuk;Chang, Jong-Soo
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.419-425
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    • 2006
  • Phospholipase $C-{\gamma}1\;(PLC-{\gamma}1)$ is an important signaling molecule for cell proliferation and differentiation. $PLC-{\gamma}1$ contains two pleckstrin homology (PH) domains, which are responsible for protein-protein interaction and protein-lipid interaction. $PLC-{\gamma}1$ also has two Src homology (SH)2 domains and a SH3 domain, which are responsible for protein- protein interaction. To identity proteins that specifically binds to PH domain of $PLC-{\gamma}1$, we prepared and incubated the glutathione S-transferase(GST)-fused PH domains of $PLC-{\gamma}1$ with COS7 cell lysate. We found that 90 kDa protein specifically binds to PH domain of $PLC-{\gamma}1$. By matrix-assisted laser desorption ionization time of flight-mass spectrometry, the 90 kDa protein revealed to be heat shock protein (Hsp) $90{\beta}$. Hsp $90{\beta}$ is a molecular chaperone that stabilizes and facilitates the folding of proteins that are involved in cell signaling, including receptors for steroids hormones and a variety of protein kinases. To know whether Hsp $90{\beta}$ affects on $PLC-{\gamma}1$ activity, we performed $PIP_2$ hydrolyzing activity of $PLC-{\gamma}1$ in the presence of purified Hsp $90{\beta}$ in vitro. Our results show that the Hsp $90{\beta}$ dose-dependently inhibits the enzymatic activity of $PLC-{\gamma}1$ and further suggest that Hsp $90{\beta}$ regulates cell growth and differentiation via regulation of $PLC-{\gamma}1$ activity.

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Rabbit Antibody Raised against Murine Cyclin D3 Protein Overexpressed in Bacterial System

  • Jun, Do-Youn;Kim, Mi-Kyung;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.474-481
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    • 1996
  • Since the commercially available rabbit anti-cyclin D3, generated from c-terminal 16 amino acid residues which are common to human and murine cyclin D3, is highly cross-reactive with many other cellular proteins of mouse, a new rabbit polyclonal anti-cyclin D3 has been raised by using murine cyclin D3 protein expressed at a high level in Escherichia coli as the immunogen. To express murine cyclin D3 protein in E. coli, the cyclin D3 cDNA fragment encoding c-terminal 236 amino acid residues obtained by polymerase chain reaction (PCR) was inserted into the NcoI/BamHI site of protein expression vector, pET 3d. Molecular mass of the cyclin D3 overexpressed in the presence of IPTG (Isopropyl $\beta$-D-thiogalactopyranoside) was approximately 26 kDa as calculated from the reading frame on the DNA sequence, and the protein was insoluble and mainly localized in the inclusion bodies that could be easily purified from the other cellular soluble proteins. When renaturation was performed following denaturation of the insoluble cyclin D3 protein in the inclusion bodies using guanidine hydrochloride, 4.4 mg of soluble form of cyclin D3 protein was produced from the transformant cultured in 100ml of LB media under the optimum conditions. Four-hundred micrograms of the soluble form of cyclin D3 protein was used for each immunization of a rabbit. When the antiserum obtained 2 weeks after tertiary immunization was applied to Western blot analysis, it was able to detect 33 kDa cyclin D3 protein in both murine lymphoma cell line BW5147.G.1.4 and human Jurkat T cells at 3,000-fold dilution with higher specificity to murine cyclin D3, demonstrating that the new rabbit polyclonal anti-murine cyclin D3 generated against c-terminal 236 amino acid residues more specifically recognizes murine cyclin D3 protein than does the commercially available rabbit polyclonal antibody raised against c-terminal 16 amino acids residues.

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Evaluation of Protein Content and Qaulity in Brown Rice by a Dye-binding Method (염색법에 의한 쌀 단백질의 함량 및 질의 평가에 관한 연구)

  • Kim, Seung-Kon;Lee, Chun-Yung;Park, Hoon
    • Korean Journal of Food Science and Technology
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    • v.3 no.2
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    • pp.101-104
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    • 1971
  • The dye-binding method based on the reaction of acidic orange-G dye with basic groups of protein molecule was investigated to observe its applicability to the determination of protein, basic amino acid and lysine contents in brown rice of several high-protein rice lines. The protein content of rice samples ranged from 7.91 to 10.53% and from 8.93 to 11.96% in terms of wet and dry bases, respectively. The correlation between dye-binding absorbance and protein content in terms of both dry and wet bases was highly significant; their correlation coefficients being $-0.955^{**}\;and\;-0.975^{**}$, respectively. The correlation of dye-binding absorbance lysine and basic amino acids were highly significant and their correlation coefficients were similar. Dye-binding absorbance-lysine showed a lower correlation than dye-binding absorbance-protein but a higher correlation than protein-lysine.

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Production of Recombinant GG1234-DsRed Fusion Protein and Its Effect on in vitro CaCO3 Crystallization (재조합 GG1234-DsRed 융합 단백질의 생산 및 In vitro 탄산칼슘 결정화에 미치는 영향에 대한 연구)

  • Son, Chaeyeon;Kim, Jin Ho;Kim, Ji Ha;Choi, Yoo Seong
    • KSBB Journal
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    • v.30 no.6
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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.