• Journal of Life Science
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• v.14 no.5
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• pp.752-760
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• 2004

The function of the [4Fe-4S] cluster containing iron (Fe-) protein in nitrogenase catalysis is to serve as the nucleotide-dependent electron donor to the MoFe protein which contains the sites for substrate binding and reduction. The ability of the Fe protein to function in this manner is dependent on its ability to adopt the appropriate conformation for productive interaction with the MoFe protein and on its ability to change redox potentials to provide the driving force required for electron transfer. The MgADP-bound (or off) conformational state of the nitrogenase Fe protein structure described reveals mechanisms for long-range communication from the nucleotide-binding sites to control affinity of association with the MoFe protein component. Two pathways, termed switches I and II, appear to be integral to this nucleotide signal transduction mechanism. In addition, the structure of the MgADP bound Fe protein provides the basis for the changes in the biophysical properties of the [4Fe-4S] observed when Fe protein binds nucleotides. The structures of the nitrogenase Fe protein with defined amino acid substitutions in the nucleotide dependent signal transduction pathways of the Switch I and Switch II have been determined by X-ray diffraction methods. These two pathways have been also implicated by site directed mutagenesis studies, structural analysis and analogies to other proteins that utilize similar nucleotide dependent signal transduction pathways. We have examined the validity of the assignment of these pathways in linking the signals generated by MgATP binding and hydrolysis to macromolecular complex formation and intermolecular electron transfer. The results provide a structural basis for the observed biophysical and biochemical properties of the Fe protein variants and interactions within the nitrogenase Fe protein-MoFe protein complex.

• Journal of Life Science
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• v.17 no.1 s.81
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• pp.18-23
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• 2007

Fe65, a neuron-specific adaptor protein, has two phosphotyrosine binding (PTB) domains. The second PTB (PTB2) domain interacts with intracellular domain fragment (AICD) of amyloid beta precursor protein (APP). Recent studies suggested that tile complex is composed of AICD and Fe65 transactivates genes that are responsible for neuronal cell death in Alzheimer's disease (AD). Therefore, a compound inhibiting the interaction between Fe65 and AICD can be a drug candidate to treat AD. However, it remains unclear how Fe65 recognizes AICD at a molecular level. Here, we report high-level production of the PTB2 domain of Fe65 in the baculovirus system. We found that the baculovirus system is an efficient method to obtain the Fe65 PTB2 domain, compared with the bacterial and mammalian expression systems. The purified recombinant protein was used for crystallization to determine its crystal structure helping to understand the molecular mechanism of Fe65-dependent signaling and to design its inhibitors.

• Applied Biological Chemistry
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• v.48 no.3
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• pp.189-200
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• 2005

Biological nitrogen fixation is an important process for academic and industrial aspects. This review will briefly compare industrial and biological nitrogen fixation and cover the characteristics of biological nitrogen fixation studied in Azotobacter vinelandii. Various organisms can carry out biological nitrogen fixation and recently the researches on the reaction mechanism were concentrated on the free-living microorganism, A. vinelandii. Nitrogen fixation, which transforms atmospheric $N_2$ into ammonia, is chemically a reduction reaction requiring electron donation. Nitrogenase, the biological nitrgen fixer, accepts electrons from biological electron donors, and transfers them to the active site, FeMo-cofactor, through $Fe_4S_4$ cluster in Fe protein and P-cluster in MoFe protein. The electron transport and the proton transport are very important processes in the nitrogenase catalysis to understand its reaction mechanism, and the interactions between FeMo-cofactor and nitrogen molecule are at the center of biological nitrogen fixation mechanism. Spectroscopic studies including protein X-ray crystallography, EPR and $M{\ddot{o}}ssbauer$, biochemical approaches including substrate and inhibitor interactions as well as site-directed mutation study, and chemical approach to synthesize the FeMo-cofactor model compounds were used for biological nitrogen fixation study. Recent research results from these area were presented, and finally, a new nitrogenase reaction mechanism will be proposed based on the various research results.

• Journal of the Korean Applied Science and Technology
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• v.34 no.3
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• pp.673-682
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• 2017

Protein hydrolysates derived from plants and animals having antioxidant, suppression of hypertension, immunodulatory, alleviation of pain, and antimicrobial activity has been known as playing important role like hormone. This study was fractioned to hydrolysis of Ogae egg white protein using the ultrafiltration. The antioxidant activity of the produced peptides was analyzed. As a result, the maximum value of DPPH radical scavenging was 1 kDa(70.83 %), hydroxy radical scavenging was 5 kDa(47.01 %), superoxide anion radical scavenging was 5 kDa(40.57 %), and $Fe^{2+}$ chelation ability was 5 kDa(29.87 %). Furthermore, the antioxidant Inhibition concentration ($IC_{50}$) of peptides was evaluated for each fraction. As a result, the maximum value of HDS was superoxide anion radical scavenging($IC_{50}$, 5.42 mg/ml). 1 kDa was $Fe^{2+}$ chelation ability($IC_{50}$, 1.67 mg/ml), 5 kDa was $Fe^{2+}$ cheating ability($IC_{50}$, 2.09 mg/ml), 10 kDa was $Fe^{2+}$ cheating ability($IC_{50}$, 2.61 mg/ml), papain was $Fe^{2+}$ cheating ability($IC_{50}$, 4.53 mg/ml). Therefore, we expect that peptides produced from Ogae egg white protein using 5 kDa fraction are useful as an antioxidant functional food ingredients.

• Korean Journal of Food Science and Technology
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• v.25 no.1
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• pp.46-51
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• 1993

The inactivating effect of iron(II)-ascorbate complex (Fe-Asc) on various phages was previously reported. This paper describes the molecular target in the phage virion attacked by Fe-Asc. The effect of Fe-Asc on protein was investigated with bovine serum albumin and the structural protein of phage J1. There were no differences in the SDS-polyacrylamide gel electrophoresis (patterns of these two proteins when either they were treated) with Fe-Asc or not. Also, there were no changes in the amino acid composition and ultraviolet spectrum of the proteins. The effects of Fe-Asc on DNA was investigated with pUC18 DNA, M13mpB DNA and ${\lambda}$ DNA as well as DNA from phage J1. Fe-Asc caused initially nicking of the subsequently form of pUC18 DNA to yield the open circular form and then subsequently the linear form. Strand breaks were also confirmed with M13mp8 DNA and ${\lambda}$ DNA as well as J1 DNA. The results indicate that the strand breaks in phage DNA could be responsible for the inactivation of phages by Fe-Asc.

• Korean Journal of Environmental Biology
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• v.20 no.3
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• pp.237-244
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• 2002

To analyze proteins related to antifungal activity, SAR01 strain was isolated from seaweed and identified as Streptomyces sp. from the result of FAME (fatty acid methyl ester) analysis. The isolated strain had antifungal activities against T species of plant pathogenic fungi. Antifungal activity deficient mutant (SAR 535) of Streptomyces sp. SAR01 was induced by gamma radiation $(^{60}Co,\;5kGy)$. By 2 D electrophoresis analysis, 6 protein spots were found in wild strain (SAR01) but these spots disappeared in mutant strain (SAR535). Among them, 5 proteins showed similarities to heat shock protein 70(HSP70), Fe-containing superoxide dismutase II (Fe- SODII), ribosome recycling factor (RRF), 10 kDa chnperonin (GroES) and inorganic pyrophosphatase (PPAse), respectively. It suggested that the above 6 proteins could be closely related to the antifungal activity of Streptomyces sp. SAR01.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.151-157
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• 1990

The maximum nitrogen fixation activity of the associative, microaerobic and acid tolerant bacteria, Azospirillum amazonense Kp1 was obtained with 0.2Kpa of $O_{2}$ and showed a reversible inhibition by the higher concentrations. Ammonium treatment caused a gradual inhibition of the activity up to 350mM. The nitrogenase systems were purified by gradient chromatography on DEAE-52 cellulose, heat treatment and preparative PAGE. The MoFe protein showed molecular weight of 210,000 including two nonidentical subunits with apparent molecular weights of 55,000 and 50,000 and an isoelectricpoint of 5.2 and contained 2, 24 and 28 atoms of Mo, Fe and acid labile S per molecule. The Fe protein revealed molecular weight of 66,000 including two types of subunits with molecular weights of 35,000 and 31,000 and an isoelectric point of 4.6, and contained 4 atoms of Fe and 6 atoms of S per molecule. The maximum specific nitrogenase activity attained 2,200 and 1,700nM $C_2H_4mg^{-1} min^{-1}$, respectively for MoFe and Fe proteins at pH7 and $35^{\circ}C$. The activity was lost after 10 and 30 days under the cold room ($4^{\circ}C$) condition for Fe and MoFe proteins, respectively.

• Bulletin of Food Technology
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• v.11 no.4
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• pp.49-60
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• 1998

Ascorbate-$Fe^3+$-EDTA에 의해 생성된 hydroxyl radical (HO.)을 이용하여 단백질과 지질의 산화적 손상을 유도하였으며, 이 손상에 대한 멜라토닌(melatonin), glutathione, $\alpha$-tocopherol의 방어효과를 분석하였다. 단백질로는 bovine serum albumin (BSA), 지질로는 phosphatidyl-choline (PC) liposome을 이용하였다. 그리고 BSA와 PC liposome의 혼합계에서도 각 항산화제의 효능을 분석하였다. 단백질의 산화적 손상은 carbonyl기의 생성 및 단백질 절단을, 지질산화는 lipid peroxidation (LPO, malondialdehyde 와 4-hydroxyalkenal) 분석을 통하여 조사하였다. 단백질의 산화는 멜라토닌과 glutathione이, 지질산화는 멜라토닌과 $\alpha$-tocopherol이 효과적으로 억제하였다. 그러나 glutathione은 지질, $\alpha$-tocopherol은 단백질의 산화를 억제하지 못하였다. BSA와 PC liposome이 동시에 존재하는 혼합물에서는 수용성과 지용성 특성을 모두 지닌 멜라토닌은 단백질, 지질 모두에 대해 산화방지효과가 있으나 수용성인 glutathione은 단백질에, 지용성인 $\alpha$-tocopherol은 지질에만 주로 효과가 있는 것으로 조사되었다.

• Journal of Aquaculture
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• v.10 no.1
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• pp.77-85
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• 1997

A 6-week feeding trial was conducted to determine the nutritional value of various dietary animal protein sources in juvenile Koran rockfish. White fish meal (WFM), flounder muscle meal (FMM), blood meal (BM), casein & gelain (CG), egg white albumin (EWA) and squid liver powder (SLP) were used as the animal protein sources, Crude protein content and available energy of the experimental diets were 50% and 15.9 kJ/g, respectively. There were significant differences among all dietary groups in weight gain (WG), feed efficiency (FE), and protein effiency ratio (PER). WFM and FFM were the best animal protein sources among the dietary groups. FMM diet had significantly higher (P<0.05) WG, FE, and PER values than those of fish fed the WFM diet. WG, FE, PER, and specific growth rate (SGR) values of fish fed BM diet were significantly higher (P<0.05) than those of fish fed EWA diet. Significant differences were found in whole body composition, hemoglobin (Hb), hepatosomatic index (HSI), and hematocrit (Ht). These results showed that low-temperature processing of lyophilized flounder muscle meal resulted in superior performance of rockfish relative to the other evaluated animal protein sources.

• The Korean Journal of Food And Nutrition
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• v.17 no.3
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• pp.294-301
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• 2004

Effects of superoxide dismutase(SOD), catalase(CAT), and such other proteins as bovine serum albumin(BSA), ovalbumin, lysozyme, and v-globulin on the autoxidation rates of L-ascorbic acid(AsA) in the absence of heavy metal ions and in the presence of Fe(III) or Cu(II) ions in water were examined. AsA was dissolved in a ultra-refined water at a concentration of 50 ${\mu}$M and 5 ${\mu}$M Fe(III) or 0.1 ${\mu}$M Cu(II) were added, and a oxygen gas was bubbled through the solution at a flow rate of 200 ml/min at 35$^{\circ}C$. The amount of remaining AsA in the reaction mixture was determined by using a UV spectrophotometer(at 265 nm). It was found that the Cu(II) at a concentration of 0.1 ${\mu}$M had a more accelerated for the autoxidation of AsA than Fe(III) at 5 ${\mu}$M. Moreover, it was confirmed that the ratio of remaining AsA was significantly larger in the presence of SOD, CAT, BSA, ovalbumin, lysozyme, and v-globulin than in the absence of proteins. The stabilization of AsA by various proteins were confirmed during the autoxidation of AsA in the presence of Fe(III) or Cu(II) in water. It was suggested that the non-enzymatic effects of SOD, CAT and some other proteins might be involves in the stabilization of AsA.