• 제목/요약/키워드: Conformational changes

검색결과 186건 처리시간 0.027초

Expression and pH-dependence of the Photosystem II Subunit S from Arabidopsis thaliana

  • Jeong, Mi-Suk;Hwang, Eun-Young;Jin, Gyoung-Ean;Park, So-Young;Zulfugarov, Ismayil S.;Moon, Yong-Hwan;Lee, Choon-Hwan;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
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    • 제31권6호
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    • pp.1479-1484
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    • 2010
  • Photosynthesis uses light energy to drive the oxidation of water at an oxygen-evolving catalytic site within photosystem II (PSII). Chlorophyll binding by the photosystem II subunit S protein, PsbS, was found to be necessary for energy-dependent quenching (qE), the major energy-dependent component of non-photochemical quenching (NPQ) in Arabidopsis thaliana. It is proposed that PsbS acts as a trigger of the conformational change that leads to the establishment of nonphotochemical quenching. However, the exact structure and function of PsbS in PSII are still unknown. Here, we clone and express the recombinant PsbS gene from Arabidopsis thaliana in E. coli and purify the resulting homogeneous protein. We used various biochemical and biophysical techniques to elucidate PsbS structure and function, including circular dichroism (CD), fluorescence, and DSC. The protein shows optimal stability at $4^{\circ}C$ and pH 7.5. The CD spectra of PsbS show that the conformational changes of the protein were strongly dependent on pH conditions. The CD curve for PsbS at pH 10.5 curve had the deepest negative peak and the peak of PsbS at pH 4.5 was the least negative. The fluorescence emission spectrum of the purified PsbS protein was also measured, and the ${\lambda}_{max}$ was found to be at 328 nm. PsbS revealed some structural changes under varying temperature and oxygen gas condition.

약물과 생체고분자 간의 상호작용(제 13보)-세파로친과 소혈청알부민의 결합에 미치는 이온강도, pH 및 온도의 영향 (Drug-Biomacromolecule Interaction (XIII)-Effect of ionic Strength, pH and Temperature on Binding of Cephalothin to Bovine Serum Albumin-)

  • 김종국;임연수;양지선;정은주
    • Journal of Pharmaceutical Investigation
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    • 제19권3호
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    • pp.163-171
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    • 1989
  • To investigate the protein binding characteristics of cephalothin, the effects of ionic strength, pH and temperature on the binding of cephalothin to bovine serum albumin (BSA) were studied by UV difference spectrophotometric method. With increasing ionic strength at constant PH and temperature, association constant decreased, but the number of binding sites sites was about 2 constantly. It may be deduced that the binding process is not only due to electrostatic forces. And the increased association constant at high ionic strength is explained by conformational changes of BSA from complex to subunits. The pH effect on the affinity of interaction indicated that the binding affinity of drug is higher in the neutral region than in the alkaline region. And, at high pH value, the number of binding sites decreased from 2 to 1 because of the conformational changes of BSA in alkaline region. The decrease in binding affinity of BSA to drug with increasing temperature was characteristic of an exothermic reaction. And the negative sign of ${\Delta}G^{\circ}$ meant that the binding process occurs spontaneously under the experimental conditions. In cephalothin-BSA complex formation, since the net enthalpy change value and entropy change value are positive, it is assumed that hydrophobic bindings are predominant in this binding process.

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Insight into Structural Aspects of Histidine 284 of Daphnia magna Arginine Kinase

  • Rao, Zhili;Kim, So Young;Li, Xiaotong;Kim, Da Som;Kim, Yong Ju;Park, Jung Hee
    • Molecules and Cells
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    • 제43권9호
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    • pp.784-792
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    • 2020
  • Arginine kinase (AK), a bioenergy-related enzyme, is distributed widely in invertebrates. The role of highly conserved histidines in AKs is still unascertained. In this study, the highly conserved histidine 284 (H284) in AK of Daphnia magna (DmAK) was replaced with alanine to elucidate the role of H284. We examined the alteration of catalytic activity and structural changes of H284A in DmAK. The catalytic activity of H284A was reduced dramatically compared to that in wild type (WT). Thus the crystal structure of H284A displayed several structural changes, including the alteration of D324, a hydrogen-bonding network around H284, and the disruption of π-stacking between the imidazole group of the H284 residue and the adenine ring of ATP. These findings suggest that such alterations might affect a conformational change of the specific loop consisting of G310-V322 at the antiparallel β-sheet region. Thus, we speculated that the H284 residue might play an important role in the conformational change of the specific loop when ATP binds to the substrate-binding site of DmAK.

A Themotropic Behavior of Egg PC Liposome Containing the Very Long Chain Fatty Acyl Component,${\alpha},{\omega}$-13,16-Dimethyloctacosanedioate Dimethyl Ester(DME C30) Isolated from The Thermophilic Anaerobic Bacteria, Thermoanaerobacter ethanolicus

  • 김현명;강세병;정승호
    • Bulletin of the Korean Chemical Society
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    • 제22권9호
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    • pp.979-983
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    • 2001
  • Thermoanaerobacter ethanolicus is a strictly anaerobic and thermophilic bacterium whose optimum temperature ranges over $65-68^{\circ}C.$ T. ethanolicus was known to contain a bipolar very long chain fatty acyl component such as $\alpha$, $\omega-1316-dimethyloctacosanedioate$, as one of the major membrane components. However, exact physiological role of this unusual component in the membrane remains unknown. Such a very long chain fatty acyl component, $\alpha$, ${\omega}-1316-dimethyloctacosanedioate$, dimethyl ester (DME C30), was isolated, and purified from the membrane of T. ethanolicus. As a function of added concentrations of the $\alpha$, $\omega-1316-dimethyloctacosanedioate$, dimethyl ester (DME C30) or cholesterol into the standard liposomes, the acyl chain ordering effect was investigated by the steady-state anisotropy with 1,6-diphenyl-1,3,5-hexatriene (DPH) as a fluorescent probe. Acyl chain order parameter (S) of vesicles containing DME C30 is higher comparing with phosphatidylcholine (PC) only vesicles. This result was discussed thermodynamically with the aid of the simulated annealing molecular dynamics simulations. Through the investigation of all the possible conformational changes of DME C30 or cholesterol, we showed that DME C30 is very flexible and its conformation is variable depending on the temperature comparing with cholesterol, which is rigid and restricted at overall temperature. We propose that the conformational change of DME C30, not the configurational change, may be involved in the regulation of the membrane fluidity against the changes of external temperature.

Structural Changes of the Spinach Photosystem II Reaction Center After Inactivation by Heat Treatment

  • Jang, Won-Cheoul;Tae, Gun-Sik
    • BMB Reports
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    • 제29권1호
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    • pp.58-62
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    • 1996
  • The structural changes in the electron donor side of the PSII reaction center have been monitored since heat treatment ($45^{\circ}C$ for 5 min) of thylakoids is known to decrease the oxygen evolving activity. In heat-treated spinach chloroplast thylakoids, the inhibitory effect of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the electron transport activity of the PSII reaction center from diphenyl carbazide to dichlorophenolindophenol became reduced approximately 3.8 times and [$^{14}C$]-labeled DCMU binding on the D1 polypeptide decreased to 25~30% that of intact thylakoid membranes, implying that the conformational changes of the DCMU binding pocket, residing on the D1 polypeptide, occur by heat treatment. The accessibility of trypsin to the $NH_2$-terminus of the cytochrome b-559 ${\alpha}$-subunit, assayed with Western blot using an antibody generated against the synthetic peptide (Arg-68 to Arg-80) of the COOH-terminal domain, was also increased, indicating that heat-treatment caused changes in the structural environments near the stromal side of the cytochrome b-559 ${\alpha}$-subunit, allowing trypsin more easily to cleave the $NH_2$-terminal domain. Therefore, the structural changes in the electron donor side of the PSII reaction center complexes could be one of the reasons why the oxygen evolving activity of the heat-treated thylakoid membranes decreased.

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Changes in Physicochemical Properties of Soybean Protein due to Acetylation during Incubation with Glucose

  • Kim, Myung-Hee;Kim, Jin-Hee;Kim, Kang-Sung
    • 한국환경보건학회지
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    • 제30권4호
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    • pp.308-313
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    • 2004
  • Native and acetylated soybean protein with acetylation percentage of $25\%$ were incubated with glucose to induce Maillard reaction. Acetylation of ${\varepsilon}$-amino group of lysine residues changed the conformation of soybean protein. The direct uv spectrum of native and acetylated soybean protein showed conformational changes with accessibility of tyrosine and tryptophan residues increased. Acetylation suppressed Maillard reaction between soybean protein and glucose. Acetylated soybean protein showed improved water sorption, fat binding, foam formation, and emulsion activity of the protein, but depressed brown pigment development and trypsin digestion. Thus aceylation prevented deterioration of certain functional characteristics that occurred during storage, besides causing functional characteristics to be improved on its own.

The Regulatory Domain of Troponin C: To Be Flexible or Not To Be Flexible

  • Gagne, Stephane M.;Sykes, Michael T.;Sykes, Brain D.
    • 한국자기공명학회논문지
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    • 제2권2호
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    • pp.131-140
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    • 1998
  • The calcium-induced structural changes in the skeletal muscle regulatory protein troponin C (NTnC) involve a transition from a ‘closed’to an ‘open’structure with the concomitant exposure of a large hydrophobic interaction site for target proteins. Structural studies have served to define this conformational change and elucidate the mechanism of the linkage between calcium binding and the induced structural changes. There are now several structures of NTnC available from both NMR and X-ray crystallography. Comparison of the calcium bound structures reveals differences in the level of opening. We have considered the concept of a flexible open state of NTnC as a possible explanation for this apparent discrepancy. We also present simulations of the closed-to-open transition which are in agreement with the flexibility concept and with experimental energetics data.

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INFLUENCE OF HUMIC SUBSTANCE (HS) ADSORPTIVE FRACTIONATION ON PYRENE PARTITIONING TO DISSOLVED AND MINERAL-ASSOCIATED HS

  • Hur, Jin;Schlautman, Mark A.
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2003년도 추계학술발표회
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    • pp.123-127
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    • 2003
  • Changes in pyrene partitioning due to mineral surface adsorptive fractionation processes of humic substances (HS) were examined in model environmental systems. For purified Aldrich humic acid(PAHA), carbon-normalized pyrene binding coefficients ( $K_{oc}$ ) for the residual (i.e., nonadsorbed and dissolved) PAHA components were different from the original dissolved PAHA $K_{oc}$ , value prior to contact with mineral suspensions. A positive correlation between the extent of pyrene binding and weight-average molecular weight (M $W_{w}$) of residual PAHA components was observed, which appeared to be unaffected by the specific mineral adsorbents use and fractionation mechanisms. A similar positive correlation was not observed with the adsorbed PAHA components, suggesting that conformational changes occurred for the mineral-associated components upon adsorption. Nonlinear pyrene sorption to mineral-associated PAHA was observed, and the degree of nonlinearity is hypothesized to be dependent on adsorptive fractionation effects and/or structural rearrangement of the adsorbed PAHA components.s.

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장력발생이 근육 분자막에 미치는 영향에 관한 연구 (A Study on the Effects of Muscle Membrane in Tension Development)

  • 김덕술;박근호
    • 한국응용과학기술학회지
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    • 제15권1호
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    • pp.47-54
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    • 1998
  • 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. The time needed to the peak tension after the onset of stimulation and the amount of peak tension depend on the number of twitch cycle. It was found that the successive twitches decreased not only the time needed to the peak tension after the onset of stimulation but also the time needed to the maximum change of the X-ray intensity. However, the difference of the time between the peak tension and the maximum intensity change($T_{i}-I_{i}$) is nearly the same at any twitch. Based on these results the causes of the decrease of $T_{i}$ and $I_{i}$, and physiological implication of $T_{i}-I_{i}$ are discussed.

Synthesis and Characterization of Polygamies and Their Metal Complexes

  • Jang, Gyu-Hwan;Kim, Yang;Lee, Man-Kil
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.346.3-346.3
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    • 2002
  • The polyamine pathway represents a logical target for chemotherapeutic intervention, since depletion of polyamines results in the disruption of a variety of cellular functions, and may in specific cases result in cytotoxicity. Polyamine interaction with DNA has also long been thought to be an important function of the natural polyamines and as more is learned about the specific interactions and the resultant conformational changes which can be influenced by the polyamine binding to DNA the potential for regional and gene-specific changes are becoming more evident. We have prepraed the elaborate polyamines by the reaction of simpler polyamines with polyalkyating agents. Synthesized polyamines were separated and purified by metal complex formation and ion-exchange chromatography. They were characterized by X-ray crystal structure determinations of their metal complexes.

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