• 제목/요약/키워드: Thermal Binding

검색결과 180건 처리시간 0.023초

Stability and Structure of S128A Mutant cAMP Receptor Protein

  • Choi, Young;Gang, JongBack
    • 통합자연과학논문집
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    • 제4권3호
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    • pp.222-226
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    • 2011
  • Cyclic AMP receptor protein(CRP) is involved in the activation of many genes corresponding to catabolite enzymes in Escherichia coli. In this study, mutant CRP(S128A) was used to elucidate the effect of Ser 128 on the cAMP-induced structural change. Based on the protease digestion and thermal analysis, serine 128 in CRP affects the cAMP binding capability and then structural change of CRP protein. In addition, CD spectra in near UV region revealed that S128A CRP retained the sensitive conformation to thermal effect relative to that of wild-type CRP, in spite of identical Tm values in the absence of cAMP.

An FCA-mediated epigenetic route towards thermal adaptation of autotrophic development in plants

  • Lee, Hyo-Jun;Ha, Jun-Ho;Park, Chung-Mo
    • BMB Reports
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    • 제50권7호
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    • pp.343-344
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    • 2017
  • Plants are able to recognize even small changes in surrounding temperatures to optimize their growth and development. At warm temperatures, plants exhibit diverse architectural adjustments, including hypocotyl and petiole elongation, leaf hyponasty, and reduced stomatal density. However, it was previously unknown how such warm temperatures affected the early stages of seedling development. In our recent study, we demonstrated that the RNA-binding protein, FCA, is critical for sustaining chlorophyll biosynthesis during early seedling development, which is a prerequisite for autotrophic transition at warm temperatures. FCA plays a dual role in this thermal response. It inhibits the rapid degradation of protochlorophyllide oxidoreductases (PORs) that mediate chlorophyll biosynthesis. In addition, it induces the expression of POR genes at the chromatin level, which contributes to maintaining functional enzyme levels. Our findings provide molecular basis for the thermal adaptation of chlorophyll biosynthesis during the early stages of seedling development in nature.

Contribution of Second Metal Binding Site for Metal Specificity of D-Xylose Isomerase

  • Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • 제9권6호
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    • pp.757-763
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    • 1999
  • The metal specificity of D-xylose isomerase from Streptomyces rubiginosus was examined by site-directed mutagenesis. The activation constants for metal ion ($Mg^{2+},{\;}Mn^{2+},{\;}or{\;}Co^{2+}$) of wild-type and mutant enzymes were determined by titrating the metal ion-free enzyme with $Mg^{2+},{\;}Mn^{2+},{\;}and{\;}Co^{2+}$, respectively. Substitutions of amino acids either on coordinated or around the M2 site (His-22O, Asn-185, Glu-186, and Glu-221) dramatically affected the activation constants as well as activity. A decrease of metal binding affinity was most significant in the presence of $Mg^{2+}$. When compared with the wild-type enzymes, the binding affinity of H220S and Nl85K for Mg^{2+} was decreased by 10-15-fold, while the affinity for $Mn^{2+}{\;}or{\;}Co^{2+}$ only decreased by 3-5-fold. All the mutations close to the M2 site changed their metal preference from $Mg^{2+}{\;}to{\;}Mn^{2+}{\;}or{\;}Co^{2+}$. These altered metal preferences may be caused by a relatively weak binding affinity of $Mg^{2+}$ to the enzyme. Thermal inactivation studies of mutants at the M2 site also support the importance of the M2 site geometry for metal specificity as well as the thermostability of the enzyme. Mutations of other important groups hardly affected the metal preference, although pronounced effects on the kinetic parameters were sometimes observed. This study proposes that the metal specificity of D-xylose isomerase can be altered by the perturbation of the M2 site geometry, and that the different metal preference of Group I and GroupII D-xylose isomerases may be caused by nonconserved amino acid residues around the M2 site.

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다중써멀버블 잉크젯방식의 3D 프린팅 시스템 개발 및 성능평가 (Evaluation and Development of Multi Thermal Bubble Ink Jet 3D Printing System)

  • 신문관;배성우;김정수
    • 한국정밀공학회지
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    • 제32권9호
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    • pp.787-792
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    • 2015
  • Recently, 3D printing technology is a hot issue in various industrial fields. According to the user's application, it allows for the free form fabrication method to be utilized in a wide range. The powder based fusion technique is one of the 3D printing methods. When using this method it is possible to apply the various binder jetting techniques such as piezo, thermal bubble jet, dispenser and so on. In this paper, a multi thermal bubble ink jet was integrated for jetting of powder binding material and developing a power fused 3D printing system. For high quality 3D printing parts, it needs an analysis and evaluation of the behavior of the thermal bubble ink jet head. In the experiment, a correlation between jetting binder quantity and layer thickness of powder was investigated, and a 3D part model was fabricated, which was used by measuring the scale factor.

SHS 합성에 의한 몰리브덴계 용사용 복합분말의 제조 (Fabrication of Mo based Thermal Spray Composite Powder by Self- propagating High- temperature Synthesis)

  • 박제신;심건주
    • 한국재료학회지
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    • 제11권9호
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    • pp.763-768
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    • 2001
  • Molybdenum-based thermal spray powder is widely used for coating the moving parts of the internal combustion engines due to its excellent wear resistance. A composite powder of the $Mo_{40}(Al_{1-x}Si_x)_{60}$ system was synthesized using the SHS method. The synthesized bulk was pulverized and specially treated to produce thermal spray powder. It was found that the synthesis reaction consisted of two-steps: the formation of $Al_8/Mo_3$ and the formation of Mo(Al,Si)$_2$. Both the temperature and the rate of the SHS reaction linearly increased with the increase of the value of x in $Mo_{40}(Al_{1-x}Si_x)_{60}$, The temperature and the rate of the reaction were also affected by the compacting density of the specimens, exhibiting the maximum valves at 62% and 60%, respectively. Since spherical shape is advantageous to the thermal spraying process, shape-control of the powder was attempted with PVA as a binding additive, resulting in the successful production of almost perfectly spherical powder of 80 $\mu\textrm{m}$ Ø$(d_{50})$ mean particle size.

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ISFET 이온감지기구의 Site Binding 모형 확장과 그 $Si_3N_4$ 수소이온 감지막에의 적용 (Extension of the Site Binding Model for Ion Sensing Mechanism of ISFET and Its Application to the Hydrogen Ion Sensing $Si_3N_4$ Membrane)

  • 서화일;권대혁;이종현;손병기
    • 대한전자공학회논문지
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    • 제25권11호
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    • pp.1358-1366
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    • 1988
  • 독립적인 nitridation step이 포함된 급속 열처리 공정을 이용하여 125-180A 두께의 이중 절연박막을 단결정 실리콘 상에 형성하였다. HCl 가스의 첨가량과 공정시간의 변화에 따른 박막 특성의 변화를 고찰하였고, 이에 따른 박막의 전기적 특성을 관찰하였다. HCl 가스의 첨가에 의해 초기의 산화막 두께의 성장은 현저하게 나타났으나, nitridation 후의 박막두께의 변화는 10A 이하로 매우 저조하였다. 이중 절연박막의 항복전압은 HCl 가스의 첨가량에 비례하여 점차 증가하였고, 절연강도는 furnace나 독립적인 nitridation step이 포함되지 않은 급속 열처리 공정으로 형성한 같은 두께의 박막에 비해 높은 것으로 분석되었다.

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Synthesis, Characterization and DNA Interaction Studies of (N,N'-Bis(5-phenylazosalicylaldehyde)-ethylenediamine) Cobalt(II) Complex

  • Sohrabi, Nasrin;Rasouli, Nahid;Kamkar, Mehdi
    • Bulletin of the Korean Chemical Society
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    • 제35권8호
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    • pp.2523-2528
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    • 2014
  • In the present study, at first, azo Schiff base ligand of (N,N'-bis(5-phenylazosalicylaldehyde)-ethylenediamine) ($H_2L$) has been synthesized by condensation reaction of 5-phenylazosalicylaldehyde and ethylenediamine in 2:1 molar ratio, respectively. Then, its cobalt complex (CoL) was synthesized by reaction of $Co(OAc)_2{\cdot}4H_2O$ with ligand ($H_2L$) in 1:1 molar ratio in ethanol solvent. This ligand and its cobalt complex containing azo functional groups were characterized using elemental analysis, $^1H$-NMR, UV-vis and IR spectroscopies. Subsequently, the interaction between native calf thymus deoxyribonucleic acid (ct-DNA) and CoL complex was investigated in 10 mM Tris/HCl buffer solution, pH = 7 using UV-vis absorption, thermal denaturation technique and viscosity measurements. From spectrophotometric titration experiments, the binding constant of CoL complex with ct-DNA was found to be $(2.4{\pm}0.2){\times}10^4M^{-1}$. The thermodynamic parameters were calculated by van't Hoff equation.The enthalpy and entropy changes were $5753.94{\pm}172.66kcal/mol$ and $43.93{\pm}1.18cal/mol{\cdot}K$ at $25^{\circ}C$, respectively. Thermal denaturation experiments represent the increasing of melting temperature of ct-DNA (about $0.93^{\circ}C$) due to binding of CoL complex. The results indicate that the process is entropy-driven and suggest that hydrophobic interactions are the main driving force for the complex formation.

Conformational Change of Escherichia coli Signal Recognition Particle Ffh Is Affected by the Functionality of Signal Peptides of Ribose-Binding Protein

  • Ahn, Taeho;Ko, Ju Hee;Cho, Eun Yi;Yun, Chul-Ho
    • Molecules and Cells
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    • 제27권6호
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    • pp.681-687
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    • 2009
  • We examined the effects of synthetic signal peptides, wild-type (WT) and export-defective mutant (MT) of ribose-binding protein, on the conformational changes of signal recognition particle 54 homologue (Ffh) in Escherichia coli. Upon interaction of Ffh with WT peptide, the intrinsic Tyr fluorescence, the transition temperature of thermal unfolding, and the GTPase activity of Ffh decreased in a peptide concentration-dependent manner, while the emission intensity of 8-anilinonaphthalene-1-sulfonic acid increased. In contrast, the secondary structure of the protein was not affected. Additionally, polarization of fluorescein-labeled WT increased upon association with Ffh. These results suggest that WT peptide induces the unfolded states of Ffh. The WT-mediated conformational change of Ffh was also revealed to be important in the interaction between SecA and Ffh. However, MT had marginal effect on these conformational changes suggesting that the in vivo functionality of signal peptide is important in the interaction with Ffh and concomitant structural change of the protein.

Evaluation of Acceptor Binding Energy of Nitrogen-Doped Zinc Oxide Thin Films Grown by Dielectric Barrier Discharge in Pulsed Laser Deposition

  • Lee, Deuk-Hee;Chun, Yoon-Soo;Lee, Sang-Yeol;Kim, Sang-Sig
    • Transactions on Electrical and Electronic Materials
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    • 제12권5호
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    • pp.200-203
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    • 2011
  • In this research, nitrogen (N)-doped zinc oxide (ZnO) thin films have been grown on a sapphire substrate by dielectric barrier discharge (DBD) in pulsed laser deposition (PLD). DBD has been used as an effective way for massive in-situ generation of N-plasma under conventional PLD process conditions. Low-temperature photoluminescence spectra of N-doped ZnO thin films provided near-band-edge emission after a thermal annealing process. The emission peak was resolved by Gaussian fitting and showed a dominant acceptor-bound excitation peak ($A^{\circ}X$) that indicated acceptor doping of ZnO with N. The acceptor binding energy of the N acceptor was estimated to be approximately 145 MeV based on the results of temperature-dependent photoluminescence (PL) measurements.

Antagonistic effects Na+ and Mg2+ on the structure, function, and stability of mycobacteriophage L1 repressor

  • Bandhu, Amitava;Ganguly, Tridib;Chanda, Palas K.;Das, Malabika;Jana, Biswanath;Chakrabarti, Gopal;Sau, Subrata
    • BMB Reports
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    • 제42권5호
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    • pp.293-298
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    • 2009
  • Temperate mycobacteriophage L1 encodes an unusual repressor (CI) for regulating its lytic-lysogenic switching and, in contrast to the repressors of most temperate phages, it binds to multiple asymmetric operator DNAs. Here, ions like $Na^+$, $Cl^-$, and $acetate^-$ ions were demonstrated to facilitate the optimal binding of CI to cognate operator DNA, whereas $K^+$, $Li^+$, ${NH_4}^+$, $Mg^{2+}$, $carbonate^{2-}$, and $citrate^{3-}$ ions significantly affected its operator binding activity. Of these ions, $Mg^{2+}$ unfolded CI most severely at room temperature and, compared to $Mg^{2+}$, $Na^+$ provided improved thermal stability to CI. Furthermore, the intrinsic tryptophan fluorescence of CI was changed notably upon replacing $Na^+$ with $Mg^{2+}$ and these opposing effects of $Mg^{2+}$ and $Na^+$ were also noticed in their actions on the C-terminal fragment (CTD) of CI. Taken together, $Na^+$ appeared to be more appropriate than $Mg^{2+}$ for maintaining the biologically active conformation of CI needed for its optimal binding to operator DNA.