• Title/Summary/Keyword: Conformational changes

검색결과 186건 처리시간 0.047초

Protein Structural Characterization by Hydrogen/Deuterium Exchange Mass Spectrometry with Top-down Electron Capture Dissociation

  • Yu, Hai Dong;Ahn, Seonghee;Kim, Byungjoo
    • Bulletin of the Korean Chemical Society
    • /
    • 제34권5호
    • /
    • pp.1401-1406
    • /
    • 2013
  • This study tested the feasibility of observing H/D exchange of intact protein by top-down electron capture dissociation (ECD) mass spectrometry for the investigation of protein structure. Ubiquitin is selected as a model system. Local structural information was obtained from the deuteration levels of c and $z^{\cdot}$ ions generated from ECD. Our results showed that ${\alpha}$-helix region has the lowest deuteration level and the C-terminal fraction containing a highly mobile tail has the highest deuteration level, which correlates well with previous X-Ray and HDX/NMR analyses. We studied site-specific H/D exchange kinetics by monitoring H/D exchange rate of several structural motives of ubiquitin. Two hydrogen bonded ${\beta}$-strands showed similar HDX rates. However, the outer ${\beta}$-strand always has higher deuteration level than the inner ${\beta}$-strand. The HDX rate of the turn structure (residues 8-11) is lower than that of ${\beta}$-strands (residues 1-7 and residues 12-17) it connects. Although isotopic distribution gets broader after H/D exchange which results in a limited number of backbone cleavage sites detected, our results demonstrate that this method can provide valuable detailed structural information of proteins. This approach should also be suitable for the structural investigation of other unknown proteins, protein conformational changes, as well as protein-protein interactions and dynamics.

Construction of 19F-13C Solid-State NMR Probe for 400MHz Wide-Bore Magnet

  • Jeong, Ji-Ho;Park, Yu-Geun;Choi, Sung-Sub;Kim, Yongae
    • 한국자기공명학회논문지
    • /
    • 제17권2호
    • /
    • pp.81-85
    • /
    • 2013
  • Various fluorine-containing materials are used in electronic devices like LCD display panels and Li-ion batteries. The structural conformation of fluorine in fluorinated materials is an important contributing factor that influences the chemical and physical properties. The conformation can be changed by heat and stress during manufacture or use. Understanding the conformational changes is critical for understanding the performance and durability of electronic devices. Solid-state NMR spectroscopy could be widely used for the analysis of various fluorine-containing materials for electronic devices. However, conventional CPMAS probes cannot be used for in-situ analysis of fluorine-containing electronic devices like LCD panels and Li-ion batteries. In this paper, we show the design, construction, and optimization of a $^{19}F-^{13}C$ double-resonance solid-state NMR probe for a 400MHz wide-bore magnet with a flat square coil for in-situ analysis of fluorine-containing electronic devices without observing fluorine background signals. This custom-built probe does not show any fluorine background signals, and can have higher efficiency for lossy samples.

Conductometric Behavior of Univalent Cation-Podand Complexes in Methanol

  • Kim, Dae-Yeon;Jung, Jong-Hwa;Chun, Jae-Sang;Lee, Shim-Sung
    • Bulletin of the Korean Chemical Society
    • /
    • 제15권11호
    • /
    • pp.967-971
    • /
    • 1994
  • The stability constants, log K of the 1 : 1 complexation for IA ions, $Ag^+$, and $Tl^+$ with a series of podands having different aromatic end-groups (I-IV) have been determined conductometrically in methanol at 25.0 $^{\circ}$C. Exceptionally the equivalent conductivity, ${\lambda}_{eq}\;of\;Li^+\;and\;Na^+$ were increased by the addition of I, because the complexed ions are less mobile than solvated ions. The order of log K values for I was $Ag^+{\gg}Tl^+>K^+>Na^+>Rb^+>Cs6+>Li^+$. The log K sequence of the podands for the certain cations was I>II>III${\geq}$IV. And every podands except IV showed the maximum selectivity for $Ag^+$ among the cations. These results were discussed in terms of the aromatic end-group effects, such as hetero-donor atoms or conformational changes by ${\pi}-{\pi}$ stacking interactions. The detailed conformations of ${\pi}-{\pi}$ stacking were also discussed by the observations of upfield shifts of some aromatic protons upon complexation from $^1H$ NMR spectra.

A Study of the Retention Behavior of Proteins in High-Performance Liquid Chromatography(II): The Effect of Salt and Temperature on Retention Behavior of Proteins in Hydrophobic Interaction Chromatography

  • Dai Woon Lee;Byung Yun Cho
    • Bulletin of the Korean Chemical Society
    • /
    • 제14권4호
    • /
    • pp.515-519
    • /
    • 1993
  • The retention behavior of proteins was investigated by using hydrophobic interaction chromatography (HIC), comparing to the results obtained in reversed-phase chromatography (RPC) described in the previous paper. A SynChropak propyl column was employed with 0.05 M phosphate buffer (pH 7.0) containing sodium sulfate. Conformational changes were recognized by examining Z values as a function of sodium sulfate concentration over a range of temperature between 5 and 65$^{\circ}C$. Z values did not change significantly at the range of the temperature showing the consistent ${\Delta}H^{\circ}$ and ${\Delta}S^{\circ}$ values. The sign and the magnitude of ${\Delta}H^{\circ}$ and ${\Delta}S^{\circ}$ of proteins in HIC were compared with those obtained in RPC. The signs of ${\Delta}H^{\circ}$ and ${\Delta}S^{\circ}$ of proteins in HIC were all positive, while those of proteins in RPC were all negative. These results suggested that the retention of proteins in HIC and in RPC were entropy-driven and enthalpy-driven process, respectively. From the two different investigations, it was concluded that the retention mechanism of RPC and HIC was based on the same fundamental principle in which separation is dependent on hydrophobicity, but the retention behavior of the proteins in HIC is clearly different from that observed in RPC.

Role of Helix 8 in Dopamine Receptor Signaling

  • Yang, Han-Sol;Sun, Ningning;Zhao, Xiaodi;Kim, Hee Ryung;Park, Hyun-Ju;Kim, Kyeong-Man;Chung, Ka Young
    • Biomolecules & Therapeutics
    • /
    • 제27권6호
    • /
    • pp.514-521
    • /
    • 2019
  • G protein-coupled receptors (GPCRs) are membrane receptors whose agonist-induced dynamic conformational changes trigger heterotrimeric G protein activation, followed by GRK-mediated phosphorylation and arrestin-mediated desensitization. Cytosolic regions of GPCRs have been studied extensively because they are direct contact sites with G proteins, GRKs, and arrestins. Among various cytosolic regions, the role of helix 8 is least understood, although a few studies have suggested that it is involved in G protein activation, receptor localization, and/or internalization. In the present study, we investigated the role of helix 8 in dopamine receptor signaling focusing on dopamine D1 receptor (D1R) and dopamine D2 receptor (D2R). D1R couples exclusively to Gs, whereas D2R couples exclusively to Gi. Bioinformatic analysis implied that the sequences of helix 8 may affect GPCR-G protein coupling selectivity; therefore, we evaluated if swapping helix 8 between D1R and D2R changed G protein selectivity. Our results suggest that helix 8 is not involved in D1R-Gs or D2R-Gi coupling selectivity. Instead, we observed that D1R with D2R helix 8 or D1R with an increased number of hydrophobic residues in helix 8 relative to wild-type showed diminished ${\beta}$-arrestin-mediated desensitization, resulting in increased Gs signaling.

Thrombus Formation Inhibition of Esculetin through Regulation of Cyclic Nucleotides on Collagen-Induced Platelets

  • Lee, Dong-Ha
    • 대한의생명과학회지
    • /
    • 제27권4호
    • /
    • pp.270-276
    • /
    • 2021
  • Physiological agents trigger a signaling process called "inside-out signaling" and activated platelets promote adhesion, granule release, and conformational changes of glycoprotein IIb/IIIa (αIIb/β3). Activated αIIb/β3 interacts with fibrinogen and initiates a second signaling step called "external signaling". These two signaling pathways can cause hemostasis or thrombosis, and thrombosis is a possible medical problem in arterial and venous vessels, and platelet-mediated thrombosis is a major cause of cardiovascular disease (CVD). Therefore, modulating platelet activity is important for platelet-mediated thrombosis and cardiovascular disease. Esculetin is a coumarin-based physiologically active 6,7-dihydroxy derivative known to have pharmacological activity against obesity, diabetes, renal failure and CVD. Although some studies have confirmed the effects of esculetin in human platelet activation and experimental mouse models, it is not clear how esculetin has antiplatelet and antithrombotic effects. We confirmed the effect and mechanism of action of escultein on human platelets induced by collagen. As a result, esculetin decreased Ca2+ recruitment through upregulation of inositol 1, 4, 5-triphosphate receptor. In addition, esculetin upregulates cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP)-dependent pathways and inhibits fibrinogen binding and thrombus contraction. Our results demonstrate the antiplatelet effect and antithrombotic effect of esculetin in human platelets. Therefore, we suggest that esculetin could be a potential phytochemical for the prevention of thrombus-mediated CVD.

Comparison Study on Changes of Antigenicities of Egg Ovalbumin Irradiated by Electron Beam or X-Ray

  • Kim, Mi-Jung;Lee, Ju-Woon;Sung, Nak-Yoon;Kim, Su-Min;Hwang, Young-Jung;Kim, Jae-Hun;Song, Beom-Seok
    • 한국축산식품학회지
    • /
    • 제34권5호
    • /
    • pp.570-575
    • /
    • 2014
  • This study was conducted to compare the effects of two forms of radiation (electron and X-ray; generated by an electron beam accelerator) on the conformation and antigenic properties of hen's egg albumin, ovalbumin (OVA), which was used as a model protein. OVA solutions (2.0 mg/mL) were individually irradiated by electron beam or X-ray at the absorbed doses of 0 (control), 2, 4, 6, 8, and 10 kGy. No differences between the two forms of radiation on the structural properties of OVA were shown by spectrometric and electrophoretic analyses. The turbidity of OVA solution increased and the main OVA bands on polyacrylamide gels disappeared after irradiation, regardless of the radiation source. In competitive indirect enzyme-linked immunosorbent assay, OVA samples irradiated by electron beam or X-ray showed different immunological responses in reactions with monoclonal and polyclonal antibodies (immunoglobulin G) produced against non-irradiated OVA. The results indicate that electron beam irradiation and X-ray irradiation produced different patterns of structural changes to the OVA molecule.

중국 중산복의 변천 및 현대패션에의 적용사례 고찰 (A Review on the Transition and Application of Modern Fashion of Chinese Mao Suit)

  • 기초;백정현;배수정
    • 복식
    • /
    • 제67권3호
    • /
    • pp.31-46
    • /
    • 2017
  • The purpose of this study is to analyse the occurrence and evolution of Mao suit which is important position in the modern costume history of China and to provide a source of design inspiration to contemporary fashion designers. In the research method, literature review and case studies was conducted in parallel. For the literature review, changes of Mao suit in each age were reviewed with reference to the related documents, Chinese costume and cultural history, prior research papers and internet resources. The case analysis was qualitatively done focusing on the silhouette, color and detail of clothes in fashion collections. The scope of the study was from 1912 to 2000. The case analysis of the Mao suit applied to the contemporary fashion was made on the applications centered around 'London Collection', 'New York Collection', 'Paris Collection', 'Milan Collection' and 'Chinese Fashion Week' from 2008 to 2015. The results found that Mao suit changed into many different forms after Sun Wen designed it for the first time in 1912. This study classified it into Phase 1(1912~1927), Phase 2(1928~1965), Phase 3(1966~1977), and Phase 4(1978~2000) with historical and political issues and conformational changes in Mao suit. The frequency analysis of the cases of the fashion collections using Mao suit from 2008 to 2015 showed an increased application of Mao suit to the western collection in New York, Paris and London in 2008 due to the impact of Beijing Olympics. However, from 2009 onwards, the frequency of the utilization of Mao suit was higher in the Chinese Fashion Week and the New York Collection. This cause is explained by the fact that the designers who inspired from Mao suit in the New York Collection are American Chinese.

cis-Diamminedichloroplatinum(II)에 의한 pBR322 DNA의 변성과 구조 변화 (cis-Diamminedichloroplatinum (II) induces denaturation and conformational changes in pBR322 DNA)

  • 구자춘;임창수;한태룡;양재명
    • Applied Biological Chemistry
    • /
    • 제33권4호
    • /
    • pp.343-348
    • /
    • 1990
  • CDDP를 처리한 pBR322 DNA로 형질변환된 대장균 LE392를 ampicillin이 포함된 한천평판배지위에 도말시켰다. Ampicillin을 함유하고 있는 평판배지위에 형성된 집락수는 13.3 ${\mu}M$의 CDDP를 처리한 뒤에는 검출되지 않을 정도로 감소하였다. CDDP를 처리한 pBR322 DNA는 외가닥 DNA에 특이성이 있는 S1 핵산분해호소에 의해 전달되었고 아가로즈 겔 전기영동상에서 이동 유형이 변했다. 이러한 결과에 의하면 CDDP가 pBR322 DNA에 반응하여 이증나선의 변성과 궁극적으로는 ampicillin 저항성 유전자를 불활성화시키는 구조변화를 일으키는 것 같다.

  • PDF

Twitch Stimulation에 의한 Myosin Heads 움직임의 시간분석 연구 (A Study on The Time For Movement of Myosin Heads by the Twitch Stimulation)

  • 김덕술;정정수;박근호
    • 한국응용과학기술학회지
    • /
    • 제21권2호
    • /
    • pp.182-189
    • /
    • 2004
  • On contraction of the muscles, marked changes in X-ray reflections are observed, suggesting that conformational changes of contractile molecules and the movement of myosin heads during muscle contraction. Time slice requires tension peak after the onset of stimulation and the height of tension peak depends on the number of twitch cycle. The muscles were stimulated by five successive stimuli at an interval of 80 ms started while the tension was still being exerted by the muscles. The intensity of $I_{11}$, $I_{10}$, $143{\AA}$ and $215{\AA}$ reflection measured with 5ms time resolution and is recorded in isometric tension. The peak height of $I_{11}$ and $143{\AA}$ intensity is changed after the onset of a stimulation $I_i$, and the length of twitch is shortened by successive twitches in the case of stimulation $T_i$. On the other hand, the peak height of In and $215{\AA}$ intensity starts to decrease at the 1st twitch and remains constant at low peak height without appreciable recovery during the contraction term. In the case of successive twitch stimulation, the myosin heads of muscle are once moved from their resting position and never returned to their initial position.