• 제목/요약/키워드: dichroism technique

검색결과 11건 처리시간 0.022초

Drug-Biomacromolecule Interaction XII: Comparative binding study of sulfaethidole to bovine serum albumin by equilibrium dialysis, fluorescence probe technique, uv difference spectrophotometry and circular dichroism

  • Kim, Chong-Kook;Chun, Yang-Sook;Lah, Woon-Lyong
    • Archives of Pharmacal Research
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    • 제12권3호
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    • pp.160-165
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    • 1989
  • Binding of sulfaethidole to bovine serum albumin was studied by equilibrium dialysis, fluorescence probe technique, uv difference spectrophotometry and circular dichroism. Equilibrium dialysis method enabled us to estimate the total number of drug binding sites of albumin molecule. For sulfaethidole, albumin had 6 primary and 40 secondary binding sites. The primary and secondary binding constants were 0.9 * 10/sup 5/ M/sup -1/ and 0.2 * 10/sup 6/ M/sup -1/, respectivitely. 1-Anilino-8-naphthalenesulfonate (ANS) and 2-(4-hydroxylbenzeneazo)- benzoic acid (HBAB) were used as the fluorescence probe and the uv spectrophotometric probe, respectively. In fluorescence probe technique, results indicated that the number of higher affinity drug binding site of albumin was 1 and the number of lower affinity drug binding sites of albumin was 3, and the primary and secondary drug binding constants for bovine serum albumin were 2.15 * 10/sup 5/M/sup -1/ and 1.04 * 10/sup 5/ M/sup -1/, respectively. In uv difference spectrophotometry, binding sites were 3 and binding constant was 1.88 * 10/sup 5/M/sup -1/. The above spectrophotometry, binding sites were 3 and binding constant was 1.88 * 10/sup 5/M/sup -1/. The above results suggest that several different methods should be used in ompensation for insufficient information about drug binding to albumin molecule given by only one method.

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진동 원편광 이색성 분광기를 사용한 키랄 에폭사이드의 광학순도 분석 (Determination of Enantiopurity of Chiral Epoxides by Vibrational Circular Dichroism Spectroscopy)

  • 이주현;이충영;김건중
    • 공업화학
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    • 제23권6호
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    • pp.577-582
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    • 2012
  • 진동 원편광 이색성(VCD; Vibrational Circular Dichroism) 분석기기를 사용한 키랄 유도체의 광학순도 분석을 수행하였다. 이 분석법을 통하여 광학이성체의 절대배위와 2% 이내의 오차 범위 내에서 %EE 값을 용이하게 측정할 수 있었다. 또한 VCD분석을 연속순환방식으로 시간 변화와 함께 수행하여 키랄화합물의 %EE 값을 측정하였다. ECH와 글리시돌 두 키랄 성분이 섞인 2성분 계에서 특별한 분리조작 없이 각 성분에 대한 농도 및 %EE 변화를 동시에 모니터링하는 것이 가능하였다. 본 연구에서 응용한 VCD 분석법은 반응 중의 반응 속도 등을 연속적으로 측정하는데 유용한 기법이며, 서로 다른 키랄 화합물이 혼합되어 있는 경우에 각각의 광학순도를 시간 변화와 함께 비파괴법으로 측정하기에 편리한 방법임을 확인하였다.

광감성 폴리비닐플루오로신나메이트의 액정 배향에 관한 연구 (Studies of Liquid Crystal Alignment on the Photosensitive Polyvinylfluorocinnamate)

  • 김동수;안원술;하기룡
    • 폴리머
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    • 제31권5호
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    • pp.393-398
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    • 2007
  • 편광 푸리에 변환 적외선 분광법(polarized fourier transform infrared spectroscopy)과 자외선 분광법(ultraviolet spectroscopy)을 이용하여 액정 디스플레이 (liquid crystal display, LCD)의 배향막으로 사용 가능한 Polyvulylfluoroclnnamate(PVCN-F) 필름의 편광 자외선(Polarized UV, PUV) 조사 및 러빙에 따른 액정의 배향 메커니즘에 관한 연구를 수행하였다. UV의 조사시간이 증가함에 따라 PVCN-F cinnamoyl group의 C=C기들의 cycloaddition 반응으로 인하여 $1638cm^{-1}$에서 나타나는 vinylene -C=C- FT-IR 흡수 피크들의 면적이 감소하고, $1712cm^{-1}$에서 나타나는 conjugated C=O 신축진동에 의한 피크가 nonconjugated C=O 신축진동 피크인 $1734cm^{-1}$로 이동함을 확인하였다. 또한 PUV가 조사된 PVCN-F 배향막을 사용하여 제조된 액정 셀에서는 조사된 PUV의 극성 방향과 수직으로 액정이 배향함을 확인하였고, 러빙 방법으로 제조된 PVCN-F 배향막을 사용한 액정 셀에서는 액정이 러빙 방향과 수평으로 배향됨을 dichroic dye(이색성 염료)의 첨가 없이 편광 FT-IR 스팩트럼을 사용하여 확인하였다.

단순 반사 방법을 이용한 NLO polymer의 전기광학 계수 측정 (Simple reflection technique for measuring the electro-optic coefficient of NLO polymer)

  • 지이권;이용산;조형준;윤국로;임종선;박광서
    • 한국광학회:학술대회논문집
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    • 한국광학회 2000년도 제11회 정기총회 및 00년 동계학술발표회 논문집
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    • pp.148-149
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    • 2000
  • 유기 고분자는 기존 무기물질의 한계성을 감안해 볼 때, 전기 및 광학적 특성이 우수하여 이에 대한 연구가 매우 활발히 진행되고 있다. 이중에서 Azo 화합물은 특별한 물리적, 광학적 성질과 그 잠재적인 응용성 때문에 계속해서 지금도 많은 관심을 끌고 있다. 그래서 Azo 화합물은 액정과 함께 비선형 광학성질을 가지는 물질에 응용될 수 있으며, 또한 광 유도현상의 이색성(dichroism)및 복굴절(birefringence) 현상 같은 특성을 나타내고 있다.$^{l)2)3)}$ 선형 광학 실험중에서 단순 입사 방법$^{4)}$ 을 이용하여 전기 광학 계수를 측정하였으며, 다시 확장 지수함수(stretched exponential)모델$^{5)}$ 로써 극화(poling)와 완화(relaxation)의 꼴을 보았다. (중략)

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Inclusion Complexation of Cyclodextrin with Prothionamide in Aqueous Solution

  • Kim, Shin-Tae;Kim, Shin-Keun
    • Journal of Pharmaceutical Investigation
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    • 제12권4호
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    • pp.132-144
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    • 1982
  • The inclusion of ${\beta}-cyclodextrin$ $({\beta}-CyD)$ with prothionamide in aqueous phase was investigated by circular dichroism(CD), ultraviolet (UV) absorption, and solubility technique. The results suggested that a region of drug chromophore was located within the asymmetric center of ${\beta}-cyclodextrin$. Solubility and spectral changes were quantitatively treated to obtain stoichiometric ratio, which was found to be 1 : 1, and formation constants which were determined by solubility, CD, and UV method were 257, 367, and 389 $M^{-1}$, respectively. Also, the formation constant of the inclusion complex was determined by CD method at various pH. The result was that $K_c$ depended upon the pH of medium, and this fact also supported that thioamide moiety was accomodated in the cavity of ${\beta}-cyclodextrin$.

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Construction of Two Metal-ion Binding Sites to Improve the 3′-5′Exonuclease Activity of Taq DNA Polymerase

  • Park, Yong-Hyun;Kim, Jong-Moon;Choi, Hye-Ja;Kim, Seog-K.;Kim, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • 제8권5호
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    • pp.471-477
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    • 1998
  • Taq DNA polymerase from Thermus aquaticus is very useful in the polymerase chain reaction. Taq DNA polymerase is classified in the pol I family, represented by E. coli DNA polymerase I. The three-dimensional structural alignment of 3'-5'exonuclease domains from the pol I family DNA polymerases explains why Taq DNA polymerase does not carry out proofreading in polymerase chain reactions. Three sequence motifs, Exo I, II, and III, must exist to carry out 3'-5'exonuclease activity for proof- reading by a 3'-5'exonuclease reaction, but these are abolished in Taq DNA polymerase. The key catalytic module in 3'-5'exonuclease is two metal ions chelated by four active-site carboxylic amino acids. Taq DNA polymerase was mutagenized to construct the catalytic module in the active site. The circular dichroism technique supported the formation of the catalytic module, and the radioactive assay showed that the 3'-5'exonuclease activity doubled in the mutant Taq DNA polymerase.

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Room Temperature Ferromagnetism on Co and Fe Doped Multi-wall Carbon Nano-tube

  • Chae, K.H.;Gautam, S.;Yu, B.Y.;Song, J.H.;Augustine, S.;Kang, J.K.;Asokan, K.
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2011년도 제40회 동계학술대회 초록집
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    • pp.171-171
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    • 2011
  • Co and Fe doped multi-wall carbon nano-tubes (MWCNTs) synthesized by microwave plasma enhanced chemical vapor deposition (PECVD) technique are investigated with synchrotron radiations at Pohang Light Source (PAL) and European Synchrotron Radiation Facility (ESRF). Near edge x-ray absorption spectroscopy (NEXAFS) measurement at C K, Co $L_{3,2}$ and Fe $L_{3,2}$-edges, and x-ray magnetic circular dichroism (XMCD) at Co and Fe $L_{3,2}$-edges have been carried at 7B1 XAS KIST and 2A MS beamline, respectively, to understand the electronic structure and responsible magnetic interactions at room temperature. X-ray absorption spectroscopy (XAS) at C K-edge shows significant p-bonding and Co and Fe L-edges proves the presence of $Co^{2+}$ and $Fe^{2+}$ in octahedral symmetry. Co and Fe doped MWCNTs show good XMCD spectra at 300K. The effect on the magnetism is also studied through swift heavy ion (SHI) radiations and magnetism is found enhanced and change in the electronic structure in Co-CNTs is investigated.

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Investigation on the Effects of Three X${\rightarrow}$Histidine Replacements on Thermostability of ${\alpha}$-Amylase from Bacillus amyloliquefaciens

  • Haghani, Karimeh;Khajeh, Khosro;Naderi-Manesh, Hossein;Ranjbar, Bijan
    • Journal of Microbiology and Biotechnology
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    • 제22권5호
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    • pp.592-599
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    • 2012
  • Bacillus licheniformis ${\alpha}$-amylase (BLA), a thermophilic counterpart of Bacillus amyloliquefaciens ${\alpha}$-amylase (BAA), is an appropriate model for the design of stabilizing mutations in BAA. BLA has 10 more histidines than BAA. Considering this prominent difference, in the present study, three out of these positions (I34, Q67, and P407; located in the thermostability determinant 1 region and Ca-III binding site of BAA) were replaced with histidine in BAA, using the site-directed mutagenesis technique. The results showed that the thermostability of P407H and Q67H mutants had increased, but no significant changes were observed in their kinetic parameters compared to that of the wild type. I34H replacement resulted in complete loss of enzyme activity. Moreover, fluorescence and circular dichroism data indicated a more rigid structure for the P407H variant compared with that of the wild-type BAA. However, the flexibility of Q67H and I34H mutants increased in comparison with that of wild-type enzyme.

Expressed Protein Ligation of 5-Enolpyruvylshikimate-3-phosphate (EPSP) Synthase: An Application to a Protein Expressed as an Inclusion Body

  • Kim, Hak-Jun;Shin, Hee-Jae;Kim, Hyun-Woo;Kang, Sung-Ho;Kim, Young-Tae
    • Bulletin of the Korean Chemical Society
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    • 제28권12호
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    • pp.2303-2309
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    • 2007
  • Expressed protein ligation (EPL) technique, joining recombinantly expressed proteins to polypeptides, has been widely adopted for addressing various biological questions and for drug discovery. However, joining two recombinant proteins together is sometimes difficult when proteins are expressed insoluble and unrefoldable, because ligation-active proteins via intein-fusion are obtainable when they are folded correctly. We overcame this limitation coexpressing target protein with additional methionine aminopeptidase (MAP) which enhances removal of the initiation methionine of recombinantly expressed protein. Our approach demonstrated that two domains of 46 kDa 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, a target of herbicide glyphosate, were successfully joined by native chemical ligation, although its C-terminal domain was expressed as an inclusion body. The intein-fused N-terminal fragment of EPSP synthase (EPSPSN, residues 1-237) was expressed and the ligation-active thioester tagged N-terminal fragment (EPSPSN-thioester) was purified using a chitin affinity chromatography and mercapto-ethanesulphonate (MESNA) as intein thiolysis reagent. Its Cterminal fragment (EPSPSC, residues Met237-238CYS-427), expressed as an inclusion body, was prepared from an additional MAP-expressing strain. Protein ligation was initiated by mixing ~1 mM of EPSPSN-thioester with ~2 mM of EPSPSCCYS (residues 238CYS-427). Also we found that addition of 2% thiophenol increased the ligation efficiency via thiol exchange. The ligation efficiency was ~85%. The ligated full-length EPSP synthase was dissolved in 6 M GdHCl and refolded. Circular dichroism (CD) and enzyme activity assay of the purified protein showed that the ligated enzyme has distinct secondary structure and ~115% specific activity compared to those of wild-type EPSP synthase. This work demonstrates rare example of EPL between two recombinantly expressed proteins and also provides hands-on protein engineering protocol for large proteins.