• Title/Summary/Keyword: Single molecule spectroscopy

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Diffusion-based determination of protein homodimerization on reconstituted membrane surfaces

  • Jepson, Tyler A.;Chung, Jean K.
    • BMB Reports
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    • v.54 no.3
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    • pp.157-163
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    • 2021
  • The transient interactions between cellular components, particularly on membrane surfaces, are critical in the proper function of many biochemical reactions. For example, many signaling pathways involve dimerization, oligomerization, or other types of clustering of signaling proteins as a key step in the signaling cascade. However, it is often experimentally challenging to directly observe and characterize the molecular mechanisms such interactions-the greatest difficulty lies in the fact that living cells have an unknown number of background processes that may or may not participate in the molecular process of interest, and as a consequence, it is usually impossible to definitively correlate an observation to a well-defined cellular mechanism. One of the experimental methods that can quantitatively capture these interactions is through membrane reconstitution, whereby a lipid bilayer is fabricated to mimic the membrane environment, and the biological components of interest are systematically introduced, without unknown background processes. This configuration allows the extensive use of fluorescence techniques, particularly fluorescence fluctuation spectroscopy and single-molecule fluorescence microscopy. In this review, we describe how the equilibrium diffusion of two proteins, K-Ras4B and the PH domain of Bruton's tyrosine kinase (Btk), on fluid lipid membranes can be used to determine the kinetics of homodimerization reactions.

Controlling Spin State of Magnetic Molecules by Oxygen Binding Studied Using Scanning Tunneling Microscopy

  • Lee, Soon-hyeong;Chang, Yun Hee;Kim, Howon;Kim, Kyung Min;Kim, Yong-Hyun;Kahng, Se-Jong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.145.1-145.1
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    • 2016
  • Binding and unbinding between molecular oxygen and metallo-porphyrin is a key process for oxygen delivery in respiration. It can be also used to control spin state of magnetic metallo-porphyrin molecules. Controlling and sensing spin states of magnetic molecules in such reactions at the single molecule level is essential for spintronic molecular device applications. Here, we demonstrate that spin states of metallo-porphyrin on surfaces can be controlled over by binding and unbinding of oxygen molecule, and be sensed using scanning tunneling microscopy and spectroscopy. Kondo localized state of metallo-porphyrin showed significant modification by the binding of oxygen molecule, implying that the spin state was changed. Our density functional theory calculation results explain the observations with the hybridization of unpaired spins in d and ${\pi}^*$ orbitals of metallo-porphyrin and oxygen, respectively. Our study opens up ways to control molecular spin state and Kondo effect by means of molecular binding and unbinding reactions on surfaces.

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Effect of Single Amino Acid Replacements on the Folding of $\alpha_1$-Antitrypsin

  • Lee, Cheolju;Yu, Myeong-Hee
    • Proceedings of the Korean Biophysical Society Conference
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    • 1998.06a
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    • pp.24-24
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    • 1998
  • The effect of stabilizing single ammo acid replacements at the sites of Phe 51, Ala 70, and Met 374 on the folding of $\alpha$$_1$-antitrypsin was investigated by fluorescence spectroscopy. The residues Phe 51 and Met 374 are located in the hydrophobic core of the molecule, B $\beta$-sheet.(omitted)

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N-type doping and band gap change of Calix adsorbed on Graphene

  • Park, Seon-Min;Yang, Se-Na;Kim, Gi-Jeong;Kim, Bong-Su;No, Gwang-Hyeon;Lee, Han-Gil
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.341-341
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    • 2010
  • Electronic structure of Calix adsorbed on epitaxial graphene (EG) was investigated using high resolution photoemission spectroscopy (HRPES). Increasing the deposition of calix molecule, we found that EG becomes n-type doping using secondary edge measurement (work function change). As we observe bonding nature of O 1s peak, we found that single O 1s peak can be clearly distinguished in the spectra indicating equivalent adsorption state. Finally, we were able to control the band gap of EG using valence band spectra as we change the amount of calix molecule. In this study, we will propose the possibility of band gap modulation of EG using calix molecule.

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Conformation of Single Stranded Poly(dA) and Its Interaction with 4',6-Diamidino-2-phenylindole

  • 정갑선;김명순;이길준;조태섭;Kim, Seog K.;이세윤
    • Bulletin of the Korean Chemical Society
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    • v.18 no.5
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    • pp.510-514
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    • 1997
  • We studied the interaction of 4',6-diamidino-2-phenylindole (DAPI) with single stranded poly(dA) using optical spectroscopic methods, including absorption, circular dichroism (CD), and fluorescence spectroscopy. The temperature-dependent conformation of poly(dA) was also investigated. The conformation of poly(dA) varied with temperature, which is explained by the stacking-destacking process of the adenine bases, resulting from the sugar conformation. The hypochromicity and red-shift in the absorption spectroscopy, the lack of CD change in the drag absorption region, and the fluorescence behavior, especially a great accessibility of the I2 quencher to the poly(dA)-bound DAPI, suggest that DAPI binds to the outside of poly(dA). The Job plot for the DAPI-poly(dA) mixture demonstrated that a stoichiometry of one DAPI molecule binds to the one phosphate of poly(dA).

Oxygen Chemisorption of NbC(111) Surface Studied by High-Resolution Electron Energy Loss and Ultraviolet Photoelectron Spectroscopy (고분해능 전자에너지손실 및 자외선광전자 분광법을 이용한 NbC(111)면의 산소흡착 연구)

  • Hwang, Yeon;Park, Soon-Ja;Aizawa, Takashi;Hayami, Wataru;Otani, Shigeki;Ishizawa, Yoshio
    • Korean Journal of Materials Research
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    • v.2 no.4
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    • pp.279-284
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    • 1992
  • Oxygen adsorption on the single crystal NbC(111) surface was studied by high-resolution electron energy loss and ultraviolet photoelectron spectroscopy. On the NbC(111) surface, oxygen molecules as well as oxygen atoms were adsorbed. Oxygen atoms were located at the 3-fold hollow site of the NbC(111) surface with the frequency of 548c$m^{-1}$. It was found that oxygen molecules had vibrational frequency of 968c$m^{-1}$which was much lower than that of the free oxygen molecule. Also the work function of the NbC(111) surface has increased by adsorption of oxygen molecule. These suggest electron tranfer from the NbC(111) substrate to the 2p${pi}_g$ substrate of the oxygen molecule.

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Atomic Force Microscopy of Asymmetric Membranes from Turtle Erythrocytes

  • Tian, Yongmei;Cai, Mingjun;Xu, Haijiao;Ding, Bohua;Hao, Xian;Jiang, Junguang;Sun, Yingchun;Wang, Hongda
    • Molecules and Cells
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    • v.37 no.8
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    • pp.592-597
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    • 2014
  • The cell membrane provides critical cellular functions that rely on its elaborate structure and organization. The structure of turtle membranes is an important part of an ongoing study of erythrocyte membranes. Using a combination of atomic force microscopy and single-molecule force spectroscopy, we characterized the turtle erythrocyte membrane structure with molecular resolution in a quasi-native state. High-resolution images both leaflets of turtle erythrocyte membranes revealed a smooth outer membrane leaflet and a protein covered inner membrane leaflet. This asymmetry was verified by single-molecule force spectroscopy, which detects numerous exposed amino groups of membrane proteins in the inner membrane leaflet but much fewer in the outer leaflet. The asymmetric membrane structure of turtle erythrocytes is consistent with the semi-mosaic model of human, chicken and fish erythrocyte membrane structure, making the semi-mosaic model more widely applicable. From the perspective of biological evolution, this result may support the universality of the semi-mosaic model.

Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

  • Chowdhury, Salina A.;Lim, Man-Ho
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.583-589
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    • 2011
  • Fluorescence correlation spectroscopy (FCS) is a sophisticated and an accurate analytical technique used to study the diffusion of molecules in a solution at the single-molecule level. FCS is strongly affected by many factors such as the stability of the excitation power, photochemical processes, mismatch between the refractive indices, and variations in the cover glass thickness. We have studied FCS near the surface of a cover glass by using rhodamine 123 as a fluorescent probe and have observed that the surface has a strong influence on the measurements. The temporal autocorrelation of FCS decays with two characteristic times when the confocal detection volume is positioned near the surface of the cover glass. As the position of the detection volume is moved away from the surface, the FCS autocorrelation becomes one-component decaying; the characteristic time of the decay is the same as the faster-decaying component in the FCS autocorrelation near the surface. This observation suggests that the faster component can be attributed to the free diffusion of the probe molecules in the solution, while the slow component has its origin from the interaction between the probe molecules and the surface. We have characterized the surface contribution to the FCS measurements near the surface by changing the position of the detection volume relative to the surface. The influence of the surface on the diffusion of the probe molecules was monitored by changing the chemical properties of the surface. The surface contribution to the temporal autocorrelation of the FCS strongly depends on the chemical nature of the surface. The hydrophobicity of the surface is a major factor determining the surface influence on the free diffusion of the probe molecules near the surface.

A Series of 3D Lanthanide Complexes Containing (La(III), Sm(III) and Gd(III)) Metal-organic Frameworks: Synthesis, Structure, Characterization and Their Luminescent Properties

  • Zhang, Huai-Min;Yang, Hao;Wu, Lan-Zhi;Song, Shuang;Yang, Li-Rong
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3777-3787
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    • 2012
  • Three kinds of 3D isomorphous and isostructural coordination polymers, namely, $\{[Ln_2(PDA)_3(H_2O)_3]{\cdot}0.25H_2O\}_{\infty}$ (Ln = La(1), Sm(2), and Gd(3)) ($PDA^{2-}$ = pyridine-2,6-dicarboxylate anion) have been synthesized under hydrothermal conditions and characterized by elemental analyses, IR spectroscopy, thermal analyses and single crystal X-ray diffraction. In these MOFs, Ln(III) centers adopt eight-coordinated and nine-coordinated with the $N_1O_7$ and $N_2O_7$ donor sets to construct distorted trianglar dodecahedron geometry and tricapped trigonal prism configurations, respectively. Based on the building block of tetranuclear homometallic $Ln_4C_4O_8$ unit (16-membered ring), 1-3 are connected into highly ordered 2D sheets via O-C-O linkers and further constructed into 3D architectures through hydrogen bonds. Crystallographic parameters suggest that the lanthanide contraction effect exist in these coordination polymers. Luminescent properties of the lanthanide-based MOFs (metal-organic frameworks) have been measured at room temperature, which reveal that they presenting ionselective characters toward certain metals, such as $Mg^{2+}$, $Cd^{2+}$ and $Pb^{2+}$ ions.

Nanoscale-NMR with Nitrogen Vacancy center spins in diamond

  • Lee, Junghyun
    • Journal of the Korean Magnetic Resonance Society
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    • v.24 no.2
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    • pp.59-65
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    • 2020
  • Nitrogen-Vacancy (NV) center in diamond has been an emerging versatile tool for quantum sensing applications. Amongst various applications, nano-scale nuclear magnetic resonance (NMR) using a single or ensemble NV centers has demonstrated promising results, opening possibility of a single molecule NMR for its chemical structural studies or multi-nuclear spin spectroscopy for quantum information science. However, there is a key challenge, which limited the spectral resolution of NMR detection using NV centers; the interrogation duration for NV-NMR detection technique has been limited by the NV sensor spin lifetime (T1 ~ 3ms), which is orders of magnitude shorter than the coherence times of nuclear spins in bulk liquid samples (T2 ~ 1s) or intrinsic 13C nuclear spins in diamond. Recent studies have shown that quantum memory technique or synchronized readout detection technique can further narrow down the spectral linewidth of NMR signal. In this short review paper, we overview basic concepts of nanoscale NMR using NV centers, and introduce further developments in high spectral resolution NV NMR studies.