• 한국식품과학회지
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• 제25권4호
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• pp.360-365
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• 1993

Phytate 함량이 다른 두 분리 대두단백(HSPI; high-phytate soy protein isolate, LSPI; low-phytate soy protein isolate)을 제조하여 pH 및 phytate 첨가량이 그들의 용해도와 소화율에 미치는 영향을 알아보았고 분리 대두단백을 용해도에 따라 분별한 후 그들의 특성을 전기영동을 통하여 알아보았다. LSPI와 HSPI 모두 분자량이 $13.6{\sim}81.1kDa$에 이르는 단백질 성분으로 이루어져 있으며 산성 pH에서 $18.0{\sim}35.0kDa$의 단백질은 phytate와 잘 결합하지 않는 특성을 가졌다. LSPI를 용해도에 다라 분별하였을 때 gliadin을 구성하는 단백질은 pH2에서 phytate와 쉽게 결합하지 않았다. 대두 단백질은 pepsin 소화율은 phytate 첨가량이 증가할수록 크게 저하되었다.

• 한국축산식품학회지
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• 제36권6호
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• pp.791-798
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• 2016

Free radicals may attack cells or tissue, leading to chronic diseases, and antioxidant consumption is potentially useful for removing free radicals. Egg proteins may be used as potential sources of antioxidant considering their ability of scavenging free radicals to apply for food or cosmetics industry. In this study, we obtained a natural antioxidant protein from fertilized eggs, which was a dietary supplement in some Asian countries. Meanwhile, antioxidant activities of these proteins were evaluated using different oxidation systems. With increasing incubation time, the antioxidant activity of these proteins increased during 15 d of incubation. The samples on day 15 were performed for isolation of antioxidant protein. The protein, named P4-1 (MW, 45 kDa), was isolated and purified by consecutive chromatographic methods. P4-1 contained 17 amino acids, which was determined by liquid chromatography-mass spectrometry and Amino Acid Analyzer. Moreover, the amino acid sequence was highly similar to that of ovalbumin. Differential scanning calorimetry showed that the denaturation temperature of P4-1 was $57.16^{\circ}C$. Furthermore, P4-1 suggested high oxygen radical-absorbance activity in ${\cdot}OH$ assays, and its antioxidant activity was stable at $30-50^{\circ}C$ in acidic and neutral pH. Thus, these results revealed that P4-1 may be a potential resource as a natural antioxidant.

• BMB Reports
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• 제37권2호
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• pp.254-259
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• 2004

The wild-type and temperature-sensitive (ts) repressor genes were cloned from the temperate mycobacteriophage L1 and its mutant L1cIts391, respectively. A sequencing analysis revealed that the $131^{st}$ proline residue of the wild-type repressor was changed to leucine in the ts mutant repressor. The 100% identity that was discovered between the two DNA regions of phages L1 and L5, carrying the same sets of genes including their repressor genes, strengthened the speculation that L1 is a minor variant of phage L5 or vice versa. A comparative analysis of the repressor proteins of different mycobacteriophages suggests that the mycobacteriophage-specific repressor proteins constitute a new family of repressors, which were possibly evolved from a common ancestor. Alignment of the mycobacteriophage-specific repressor proteins showed at least 7 blocks (designated I-VII) that carried 3-8 identical amino acid residues. The amino acid residues of blocks V, VI, and some residues downstream to block VI are crucial for the function of the L1 (or L5) repressor. Blocks I and II possibly form the turn and helix 2 regions of the HTH motif of the repressor. Block IV in the L1 repressor is part of the most charged region encompassing amino acid residues 72-92, which flanks the putative N-terminal basic (residues 1-71) and C-terminal acidic (residues 93-183) domains of L1 repressor.

• Toxicological Research
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• 제14권4호
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• pp.525-533
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• 1998

The covalent interactions of toluenediisocyanate (TDI) with macromolecules were investigated both in vitro and in vivo. In vitro incubations of 2,4- and 2,6-TDI with DNA or proteins resulted in dose-dependent formation of TDI-protein and TDI-DNA adducts. TDI-treated DNA was highly resistant to enzymatic digestion and thermal hydrolysis, but was readily hydrolyzed under acidic conditions by releasing its corresponding toluenediamine (TDA), suggesting that TDI caused the crosslinking of DNA. Reaction of TDI with albumin and globin resulted in the formation of several adducts, and some adducts were formed in blood of TDI-treated rats in a dose-dependent fashion. Administration of TDI to rats resulted also in a dose-dependent binding of TDI to hepatic tissue. Levels of TDI-albumin adducts were 10 times higher than those of TDI-globin adducts; the biological half lives of TDI-albumin and TDI-globin adducts were 1.2 and 12.5 days, respectively. Globin adducts were detected up to 28 days after the treatment. Hepatic TDI protein adducts were persistent for a substantial period whereas the levels of hepatic TDI-DNA adduct were decreased rapidly. These results indicate that the isocyanato group of TDI is not readily hydrolyzed under physiological conditions, is transported to other organs, and is bound to DNA and/or proteins without further metabolic activation. As the adducted products degrade in the body, TDA is released and introduced to the liver. TDA may additionally bind to hepatic tissue after metabolic activation. Thus, the toxic effect of TDI exposure is considered to persist during the lifetime of the adducted biological macromolecules.

• Journal of Microbiology and Biotechnology
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• 제34권3호
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• pp.525-537
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• 2024

Pilins are protein subunits of pili. The pilins of type IV pili (T4P) in pathogenic bacteria are well characterized, but anything is known about the T4P proteins in acidophilic chemolithoautotrophic microorganisms such as the genus Acidithiobacillus. The interest in T4P of A. thiooxidans is because of their possible role in cell recruitment and bacterial aggregation on the surface of minerals during biooxidation of sulfide minerals. In this study we present a successful ad hoc methodology for the heterologous expression and purification of extracellular proteins such as the minor pilin PilV of the T4P of A. thiooxidans, a pilin exposed to extreme conditions of acidity and high oxidation-reduction potentials, and that interact with metal sulfides in an environment rich in dissolved minerals. Once obtained, the model structure of A. thiooxidans PilV revealed the core basic architecture of T4P pilins. Because of the acidophilic condition, we carried out in silico characterization of the protonation status of acidic and basic residues of PilV in order to calculate the ionization state at specific pH values and evaluated their pH stability. Further biophysical characterization was done using UV-visible and fluorescence spectroscopy and the results showed that PilV remains soluble and stable even after exposure to significant changes of pH. PilV has a unique amino acid composition that exhibits acid stability, with significant biotechnology implications such as biooxidation of sulfide minerals. The biophysics profiles of PilV open new paradigms about resilient proteins and stimulate the study of other pilins from extremophiles.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2002년도 제38회 학술심포지움
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• pp.20-47
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• 2002

Two-dimensional gel electrophoresis (2-DE) maps for human stomach tissue proteins have been prepared by displaying the protein components of the tissue by 2-DE and identifying them using mass spectrometry. This will enable us to present an overview of the proteins expressed In human stomach tissues and lays the basis for subsequent comparative proteome analysis studies with gastric diseases such as gastric cancer. In this study, 2-DE maps of soluble fraction proteins were prepared on two gel images with partially overlapping pH ranges of 4-7 and 6-9. On the gels covering pH 4-7 and pH 6-9, about 900 and 600 protein spots were detected on silver staining, respectively. For protein identification, proteins spots on micropreparative gels stained by colloidal Coomassie Brilliant Blue G-250 were excised, digested in-gel with trypsln, and analyzed by peptide mass fingerprinting with delayed extraction-matrix assisted laser dosorption/ionization-mass spectrometry (DE-MALDI-MS). In all, 243 protein spots (168 spots in acidic map and 75 spots in basic map) corresponding to 136 different proteins were identified. Besides these principal maps, maps of lower resolution, i.e. overview maps (displayed on pH 3-10 gels) for total homogenate and soluble fraction, are also presented with some identifications mapped on them. Based on the 2-DE maps presented in this study, a 2-DE database for human stomach tissue proteome has been constructed and available at http://proteome.gsnu.ac.kr/DB/2DPAGE/Stomach/. The 2-DE maps and the database resulting from this study will serve important resources for subsequent proteomic studies for analyzing the normal protein variability in healthy tissues and specific protein variations in diseased tissues.

• Molecules and Cells
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• 제37권3호
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• pp.264-269
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• 2014

The molecular interaction between tumor suppressor p53 and the anti-apoptotic Bcl-2 family proteins plays an essential role in the transcription-independent apoptotic pathway of p53. In this study, we investigated the binding of p53 DNA-binding domain (p53DBD) with the anti-apoptotic Bcl-2 family proteins, Bcl-w, Mcl-1, and Bcl-2, using GST pull-down assay and NMR spectroscopy. The GST pull-down assays and NMR experiments demonstrated the direct binding of the p53DBD with Bcl-w, Mcl-1, and Bcl-2. Further, NMR chemical shift perturbation data showed that Bcl-w and Mcl-1 bind to the positively charged DNA-binding surface of p53DBD. Noticeably, the refined structural models of the complexes between p53DBD and Bcl-w, Mcl-1, and Bcl-2 showed that the binding mode of p53DBD is highly conserved among the anti-apoptotic Bcl-2 family proteins. Furthermore, the chemical shift perturbations on Bcl-w, Mcl-1, and Bcl-2 induced by p53DBD binding occurred not only at the p53DBD-binding acidic region but also at the BH3 peptide-binding pocket, which suggests an allosteric conformational change similar to that observed in Bcl-$X_L$. Taken altogether, our results revealed a structural basis for a conserved binding mechanism between p53DBD and the anti-apoptotic Bcl-2 family proteins, which shed light on to the molecular understanding of the transcription-independent apoptosis pathway of p53.

• 생명과학회지
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• 제27권7호
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• pp.826-833
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• 2017

대기의 이산화탄소의 농도 증가는 해양산성화와 지구온난화를 유발하는 것으로 알려져 있다. 해마는 해양생태계 및 수산자원생물로서 중요한 종으로 알려져 있지만, 최근 해양산성화로 인하여 개체수가 감소되고 있는 실정이다. 따라서 본 연구에서는 멸종 위기 종인 복해마(Hippocampus kuda)에 미치는 생리적 영향을 조사하기 위해서 사육수의 산성조건인 pH 6.0, 6.5, 7.0 및 자연해수(pH 8.0)의 환경에서 복해마(H. kuda)를 15일 동안 사육 후 체내 조성 변화 및 항산화 효소 활성 변화에 대하여 조사를 실시하였다. 복해마(H. kuda)의 크기 및 성장은 대조군인 pH 8.0을 제외한 실험군에서는 pH가 저하함에 따라 감소하는 경향을 나타내었다. 체내 조성성분인 회분, 조지방 및 조단백 또한 pH 저하에 따라 농도의존적으로 감소하는 것이 관찰되었다. SOD, CAT 및 GSH와 같은 항산화 효소의 분석 결과, SOD활성의 경우, pH 저하에 따라 농도의존적으로 감소하지만, 이와 상반되게 CAT 및 GSH에서는 pH저하에 따라 활성이 농도의존적으로 증가하는 결과가 나타내었다. 이것은 복해마(H. kuda)가 사육수의 pH 저하에 따른 체내 항상성을 유지하는 과정 중 스트레스가 야기되어 에너지 대사가 손상된 것으로 추정된다. 항산화효소는 일반적으로 산성화 스트레스에 민감하게 작용하는데 본 연구에서도 사육수의 pH 변화에 따라 항산화 효소작용이 유의하게 변화하였다. 이러한 결과로 복해마(H. kuda)에 있어서 산성화 노출을 통한 생리학적 스트레스가 항산화 반응 및 체내 성분과 성장을 저해하는 것으로 여겨진다.

• BMB Reports
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• 제33권4호
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• pp.300-307
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• 2000

Two glycoproteins were purified and biochemically characterized from the lichen X. parietina. Both behaved as enzymes with arginase activity and haemaglutinins. Secreted arginase (SA) contained galactose and glucose in the saccharide moiety and an isoelectric point of 4.54. The algal binding-protein (ABP) had N-acetyl-glucosamine and glucose as glycosidic residues and an isoelectric point of 3.53. Both proteins had the same molecular mass (58.6 kDa) and the same qualitative amino acidic composition. The results allowed us to consider these glycoproteins as isolectins, which have significant physiological roles in the relationship between photobiont and mycobiont of symbiotic association.

• 펄프종이기술
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• 제36권1호
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• pp.24-29
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• 2004

Alkaline cellulolytic enzymes from cultured medium of Coprinus cinereus 2249 were purified with gel and ion-exchange chromatography and characteristics of those enzyme proteins were investigated. A fiber length distribution and a crystallinity of cellulose and sugar composition of enzyme treated Mixed Office Wastepaper(MOW) and Unbleached Kraft Pulp(UKP) were analysed. The conclusion could summarized as follows; \circled1 Alkaline and acidic, endo- and exo-glucanases were purified from cultured medium of Coprinus cinereus 2249. \circled2 The approximate molecular weight of alkaline endo-glucanase was 42 kDa, and also that of alkaline exo-glucanase was 50 kDa. A fiber length distribution and a crystallization of cellulose and sugar composition of enzyme treated MOW and UKP were not so much changed with original paper and pulp.