• 제목/요약/키워드: Fe/S cluster

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Backbone NMR chemical shift assignment for the substrate binding domain of Escherichia coli HscA

  • Jin Hae Kim
    • 한국자기공명학회논문지
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    • 제28권2호
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    • pp.6-9
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    • 2024
  • HscA is a Hsp70-type chaperone protein that plays an essential role to mediate the iron-sulfur (Fe-S) cluster biogenesis mechanism in Escherichia coli. Like other Hsp70 chaperones, HscA is composed of two domains: the nucleotide binding domain (NBD), which can hydrolyze ATP and use its chemical energy to facilitate the Fe-S cluster transfer process, and the substrate binding domain (SBD), which directly interacts with the substrate, IscU, the scaffold protein of an Fe-S cluster. In the present work, we prepared the isolated SBD construct of HscA (HscA(SBD)) and conducted the solution-state nuclear magnetic resonance (NMR) experiments to have its backbone chemical shift assignment information. Due to low spectral quality of HscA(SBD), we obtained all the NMR data from the sample containing the peptide LPPVKIHC, the HscA-interaction motif of IscU, from which the chemical shift assignment could be done successfully. We expect that this information provides an important basis to execute detailed structural characterization of HscA and appreciate its interaction with IscU.

High-pressure NMR analysis on Escherichia coli IscU

  • Jongbum Na;Jinbeom Si;Jin Hae Kim
    • 한국자기공명학회논문지
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    • 제28권1호
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    • pp.1-5
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    • 2024
  • IscU, the iron-sulfur (Fe-S) cluster scaffold protein, is an essential protein for biogenesis of Fe-S clusters. Previous studies showed that IscU manifests a metamorphic structural feature; at least two structural states, namely the structured state (S-state) and the disordered state (D-state), interconverting in a physiological condition, was observed. Moreover, subsequent studies demonstrated that the metamorphic flexibility of IscU is important for its Fe-S cluster assembly activity as well as for an efficient interaction with various partner proteins. Although solution nuclear magnetic resonance (NMR) spectroscopy has been a useful tool to investigate this protein, the detailed molecular mechanism that sustains the structural heterogeneity of IscU is still unclear. To tackle this issue, we applied a high-pressure NMR (HP-NMR) technique to the IscU variant, IscU(I8K), which shows an increased population of the S-state. We found that the equilibrium between the S- and D-state was significantly perturbed by pressure application, and the specific regions of IscU exhibited more sensitivity to pressure than the other regions. Our results provide novel insights to appreciate the dynamic behaviors of IscU and the related versatile functionality.

Azotobacter vinelandii에서의 생물학적 질소고정 작용 메카니즘 (Mechanism of Biological Nitrogen Fixation in Azotobacter vinelandii)

  • 김용웅;한재홍
    • Applied Biological Chemistry
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    • 제48권3호
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    • pp.189-200
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    • 2005
  • 생물학적 질소고정과정의 연구는 학문적으로나 산업적으로 매우 중요한 과정이다. 본 총설에서는 공업적 질소고정과 비교되는 생물학적 질소고정의 특징을 간단히 살펴보고, Azotobacter vinelandii에서 연구되고 있는 생물학적 질소고정효소의 특징을 다룬다. 생물학적 질소고정과정은 다양한 생명체에서 일어나며, 최근에는 미생물인 A. vinelandii에 그 작용 메커니즘에 관한 연구가 집중되어 있다. 공기중의 질소를 암모니아로 변환시키는 질소고정은 화학적으로 환원 반응에 해당하므로 전자의 공급이 필요하다. 생물학적 질소고정을 담당하는 질소고정효소는 촉매반응을 위해 생물학적인 환원력을 사용하여 전자를 공급받아, Fe 단백질의 $Fe_4S_4$ cluster와 MoFe 단백질의 P-cluster를 거쳐 질소 환원 반응이 일어나는 FeMo-cofactor로 전달한다. 이러한 전자전달의 과정과 수소이온의 전달 과정은 질소고정효소의 반응 메커니즘 이해에 매우 중요한 과정이며, FeMo-cofactor와 질소분자의 상호작용은 생물학적 질소고정 메커니즘의 중심에 있다. 질소고정 작용 메커니즘의 연구에는 X-선 단백질 결정학, EPR과 $M{\ddot{u}}ssbauer$ 등의 다양한 분광학적 방법과, 효소의 기질과 저해제의 상호작용을 연구하고 mutant와 비교하는 생화학적 접근방법, 그리고 FeMo-cofactor의 모델 화합물을 합성하여 연구하는 화학적 방법 등이 적용되었다. 이들 분야의 최근 연구결과를 소개하며, 마지막으로, 다양한 연구 결과에 바탕하여 새로운 질소고정효소의 작용 기작이 제안하였다.

Mitochondrial Protein Nfu1 Influences Homeostasis of Essential Metals in the Human Fungal Pathogen Cryptococcus neoformans

  • Kim, Jeongmi;Park, Minji;Do, Eunsoo;Jung, Won Hee
    • Mycobiology
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    • 제42권4호
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    • pp.427-431
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    • 2014
  • Mitochondrial protein Nfu1 plays an important role in the assembly of mitochondrial Fe-S clusters and intracellular iron homeostasis in the model yeast Saccharomyces cerevisiae. In this study, we identified the Nfu1 ortholog in the human fungal pathogen Cryptococcus neoformans. Our data showed that C. neoformans Nfu1 localized in the mitochondria and influenced homeostasis of essential metals such as iron, copper and manganese. Marked growth defects were observed in the mutant lacking NFU1, which suggests a critical role of Nfu1 in Fe-S cluster biosynthesis and intracellular metal homeostasis in C. neoformans.

One-dimensionally Ordered Array of Co and Fe Nanoclusters on Carburized-W(110) via Template Assisted Self-Assembly

  • Kim, Ji-Hyun;Yang, Serlun;Kim, Jae-Sung;Lukashev, Pavel;Rojas, Geoffrey;Enders, Axel;Sessi, Violetta;Honolka, Jan
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2012년도 제42회 동계 정기 학술대회 초록집
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    • pp.135-136
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    • 2012
  • Carbon atoms near the surface of W(110) induce reconstructions such as $R(15{\times}12)$ -C/W(110) which consists of two characteristic parts, one square shaped and bright protrusion and two smaller ones. In the atomic resolution STM image, the bigger protrusion shows the periodicities of clean W(110), indicating that it is almost carbon poor region. The smaller protrusion contains hexagonal carbide surface layer of ${\alpha}$-W2C on W(110). Employing this carburized W(110) as templates, we grow Co and Fe clusters of less than ten atoms. Due to the selectivity of bonding sites, growth of larger cluster is highly unfavorable for Co and the size of clusters is very uniform. Since Co atoms prefer to sit on the bigger protrusion rather than smaller one, Co cluster can be arranged one-dimensionally in $R(15{\times}12)$-C/W(110) with quite uniform size distribution. However, Fe clusters sit on both sites without favored site, but still with uniform size distribution. On the other hand, Fe clusters can be grown with quasi one-dimensional order in $R(15{\times}3)$-C/W(110), which consists of only smaller protrusions. We investigate the magnetic properties of the ordered nano-sized clusters. Experiments using XMCD reveals little magnetic moment of Co cluster on $R(15{\times}12)$-C/W(110). This observation is consistent with the predictions of our first principles calculations that small Co clusters can be nonmagnetic or antiferromagnetic with low mean magnetic moment per atom.

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Isolation and Partial Characterization of Two Ferredoxins from the Photosynthetic Bacterium Heliobacillus mobilis

  • Hatano, Atsushi;Inoue, Kazuhito;Deo, Daisuke;Sakurai, Hidehiro
    • Journal of Photoscience
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    • 제9권2호
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    • pp.388-390
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    • 2002
  • Two ferredoxin (Fd) fractions, namely, Fd-A and Fd-B were isolated from Heliobacillus mobilis cells, and purified by ammonium sulfate fractionation, DEAE, gel-permeation and Phenyl-Superose column chromatographies under anaerobic conditions. Their absorption spectra were typical of 2[4Fe-4S] cluster type Fds with peaks at about 385 and 280 nm and a shoulder at about 305 nm. Their N-terminal amino acid sequences were determined, which showed that both of them contain a [4Fe-4S] cluster binding motif. Fd-B was sensitive to oxygen, and itsA$_{385}$ value decreased by about 50% in 2 h at 4$^{\circ}C$ under aerobic conditions. In contrast, $A_{385}$ of Fd-A was essentially unchanged up to 24 h under the same conditions.

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Electronic state calculation of ceramics by $DV-X\;{\alpha}$ cluster method

  • Adachi, Hirohiko
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 1994년도 추계 학술발표 강연 및 논문 개요집
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    • pp.1-1
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    • 1994
  • ;The electronic state calculations for various types of ceramic materials have beell performed by the use of $DV-X\;{\alpha}$ cluster method. The molecular orbital levels and wave functions for model clusters have been computed to study the electronic properties ami chemical bonding of the ceramics. For ${\beta}-sialon(Si_{6-z}Al_zO_zN_{8-z})$ which is a high temperature structural material based on ${\beta}-Si_3N_4$, we have made model cluster calculations to estimate the strength of chemical bonding between atoms by the Mulliken population analysis. It is found that the covalent bonding between Si and N atoms is very strong in pure ${\beta}-Si_3N_4$, but the covalency around solute atom is considerably weakened when Si atom is substituted by AI. This tendency is enhanced by an additional substitution of oxygen atom for N. The result calculated can well explain the experimental data of changes in mechanical properties such as the reductions of Young's modulus and Vickers hardness with increment of z-value in ${\beta}-sialon$. Various model clusters for transition metal oxides which show many interesting physical and chemical properties have also been calculated. High-valent perovskite-type iron oxides EMFe0_3E(M=Ca and Sr) possess very interesting magnetic and chemical properties. In these oxides, iron exists as $Fe^{4+}$ state, but the experimental measurement of Mossba~er effect suggests that disproportionation $2Fe^{4+}=Fe^{3+}+Fe^{5+}$ takes place for $CaFe0_3$ at low temperatures. The model cluster calculations for these compounds indicated the existence of considerably strong covalent bonding of Fe-O. The calculations of hyperfine interaction at iron neucleus show very good agreement with the experimental Mossbauer measurements. The result calculated also implies that the disproportionation reaction is strongly possible by assuming the quenching of breathing phonon mode at low temperatures.tures.

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MgADP 결합 및 아미노산 치환 Nitrogenase Fe 단백질의 구조 및 기능 분석 (Structural and Functional Analysis of Nitrogenase Fe Protein with MgADP bound and Amino Acid Substitutions)

  • Jeong, Mi-Suk;Jang, Se-Bok
    • 생명과학회지
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    • 제14권5호
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    • pp.752-760
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    • 2004
  • Nitrogenase 촉매에서 Fe-단백질을 포함하는 [4Fe-4S] 클라스터의 기능은 기질의 결합과 환원 자리를 포함하는 MoFe-단백질로 핵산 의존 전자 주개로 작용하는 것이다. 이러한 방법의 Fe-단백질의 기능은 Mofe-단백질과 상호작용을 위해 적합한 구조를 갖추며 전자 전달을 위한 추진력을 제공하기 위해 산화 환원 퍼텐셜을 변화시키는 능력에 의존한다. Nitrogenase Fe-단백질에 MgADP가 결합한 (혹은 떨어진) 구조적 정보는 핵산 결합 자리로부터 MoFe-단백질과의 결합력을 조절하기 위한 장거리 상호작용 메커니즘을 제시한다. 스위치 I과 II의 두 가지 경로가 뉴클레오티드의 신호전달 메커니즘을 담당한다. MgADP가 결합된 Fe-단백질의 구조는 Fe 단백질이 핵산과 결합할 때 관찰되는 [4Fe-4S] 클라스터의 생물리학적 특성 변화의 기초를 제공한다. 스위치, I과 II의 핵산 의존 신호전달 경로에서 특정 아미노산이 치환된 nitrogenase Fe-단백질의 구조들이 X-선 회절법에 의해서 결정되었다. 이들 경로는 아미노산 치환 연구, 구조 분석, 유사한 핵산 의존 신호전달 경로에 이용된 다른 단백질 등에 의해서도 분석되었다. 이들 경로가 거대분자 착물 형성과 분자간 전자 전달을 위한 MgADP 결합과 가수분해의 신호전달 경로로의 타당성이 조사되었다. 이러한 결과는 nitrogenase Fe 단백질과 MoFe-단백질 착물에서 Fe-단백질의 변이와 상호작용의 생물리학적 및 생화학적 특성을 위한 기초적 자료를 제공할 것이다.

Structures of SUF Machinery Proteins and their Implications for Iron-Sulfur Cluster Biosynthesis

  • Wada, Kei;Hasegawa, Yuko;Kitaoka, Shintaro;Takahashi, Yasuhiro;Fukuyama, Keiichi
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2006년도 International Meeting of the Microbiological Society of Korea
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    • pp.66-68
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    • 2006
  • SUF machinery in Echerichia coli, responsible for the biosynthesis of iron-sulfur clusters, is composed of six protein components (SufABCDSE), among which SufB, SufC, and SufD associate in a complex. We have determined the structures of SufA, SufC, and SufD by X-ray crystallography. SufA is a dimer, in which C-terminal segments containing essential cysteine residues (Cys-Gly-Cys) are positioned to allow coordination of an Fe-S cluster and/or an Fe atom. SufC has the overall structure similar to that of ABC-ATPase but takes an inactive form. SufD has a ${\beta}-helix$ flanked with a-helical domains. We also studied the functional roles of the residues in SufD by mutagenesis and determined the crystal structure of SufCD complex. Molecular mechanism of Fe-S cluster biosynthesis is discussed on the basis of the structural and functional evidence.

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NEAR-INFRARED PHOTOMETRIC STUDY OF THE GALACTIC OPEN CLUSTERS NGC 1641 AND NGC 2394 BASED ON 2MASS DATA

  • Kim, Sang-Chul
    • 천문학회지
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    • 제39권4호
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    • pp.115-123
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    • 2006
  • We present $JHK_S$ near-infrared CCD photometric study for the Galactic open clusters NGC 1641 and NGC 2394. These clusters have never been studied before, and we provide, for the first time the cluster parameters; reddening, distance, metallicity and age. NGC 1641 is an old open cluster with age $1.6{\pm}0.2$ Gyr, metallicity $[Fe/H]=0.0{\pm}0.2$ dex, distance modulus$(m-M)_0=10.4{\pm}0.3\;mag(d=1.2{\pm}02\;kpc)$, and reddening $E(B-V)=0.10{\pm}0.05$ mag. The parameters for the other old open cluster NGC 2394 are estimated to be $age=1.1{\pm}0.2$ Gyr, $[Fe/H]=0.0{\pm}0.2$ dex, $(m-M)_0=9.1{\pm}0.4\;mag(d=660{\pm}120\;pc)$, and $E(B-V)=0.05{\pm}0.10$ mag. The metallicities and distance values for these two old open clusters are consistent with the relation between the metallicities and the Galactocentric distances of other old open clusters. We find the metallicity gradient of 53 old open clusters including NGC 1641 and NGC 2394 to be ${\Delta}[Fe/H]/{\Delta}R_{gc}=-0.067{\pm}0.009\;dex\;kpc^{-1}$.