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Functional Diversity of Cysteine Residues in Proteins and Unique Features of Catalytic Redox-active Cysteines in Thiol Oxidoreductases

  • Fomenko, Dmitri E.;Marino, Stefano M.;Gladyshev, Vadim N.
    • Molecules and Cells
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    • v.26 no.3
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    • pp.228-235
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    • 2008
  • Thiol-dependent redox systems are involved in regulation of diverse biological processes, such as response to stress, signal transduction, and protein folding. The thiol-based redox control is provided by mechanistically similar, but structurally distinct families of enzymes known as thiol oxidoreductases. Many such enzymes have been characterized, but identities and functions of the entire sets of thiol oxidoreductases in organisms are not known. Extreme sequence and structural divergence makes identification of these proteins difficult. Thiol oxidoreductases contain a redox-active cysteine residue, or its functional analog selenocysteine, in their active sites. Here, we describe computational methods for in silico prediction of thiol oxidoreductases in nucleotide and protein sequence databases and identification of their redox-active cysteines. We discuss different functional categories of cysteine residues, describe methods for discrimination between catalytic and noncatalytic and between redox and non-redox cysteine residues and highlight unique properties of the redox-active cysteines based on evolutionary conservation, secondary and three-dimensional structures, and sporadic replacement of cysteines with catalytically superior selenocysteine residues.

Binding Studies of Erythromycin A and its Analogues using Molecular Docking Technique

  • Kamarulzaman, Ezatul Ezleen;Mordi, Mohd Nizam;Mansur, Shariff Mahsufi;Wahab, Habibah
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.35-40
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    • 2005
  • Interaction of twelve erythromycin A analogues with 50S ribosomal subunit were studied employing AutoDock 3.0.5. Results showed that all active macrolides bound at the same binding site with erythromycin A in contrast to the inactive analogues which bound at location slightly different than erythromycin A. The binding site showed consistency with the X-ray data from the perspectives of hydrogen bonding and hydrophobic interactions formed by erythromycins, roxithromycin, azithromycin, cethromycin and telithromycin with the ribosome. The inactive derivatives of erythromycin A anhydride showed higher binding free energy, while 5-desosaminyl erythronolides A and B even though having quiet similar values of binding free energy with the active analogues, docked at binding sites which are quiet different than the active analogues. These results suggest the molecular docking technique can be used in predicting the binding of erythromycin A analogues to their ribosomal target.

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Bacillus subtilis 유래 Glycerol-3-phosphate Cytidylyltransferase의 화학적 수식

  • 박영서
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.173-177
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    • 1997
  • Glycerol-3-phosphate cytidylyltransferase from Bacillus subtilis was modified with various chemical modifiers to determine the active sites of the enzyme. Treatment of the enzyme with group-specific reagents diethylpyrocarbonate, N-bromosuccinimide, or carbodiimide resulted in complete loss of enzyme activity, which shows histidine, tryptophan, and glutamic acid or aspartic acid residues are at or near the active site. In each case, inactivation followed pseudo first-order kinetics. Inclusion of glycerol-3-phosphate and/or CTP prevented the inactivation, indicating the presence of tryptophan and glutamic acid or aspartic acid residues at the substrate binding site. Analysis of kinetics of inactivation showed that the loss of enzyme activity was due to modification of a two histidine residues, single tryptophan residue, and two glutamic acid or aspartic acid residues.

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A Study on a Plan for an Active Use of a Trade e-Marketplace (무역 e-마켓플레이스의 활성화방안에 관한 연구)

  • Kim, Sun-Chul;Lee, Kil-Nam
    • International Commerce and Information Review
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    • v.9 no.1
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    • pp.187-209
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    • 2007
  • Since mid-1990, the global trade environments has been rapidly changing in many sectors due to the advancement of information technology. Of which an international trade e-Marketplace is one that has been playing a crucial role. The international trade e-Marketplace has the potential to accelerate existing trends and new ways of conducting trade. Nevertheless, e-Marketplace seems to have not been facilitated because many traders do not fully understand about performance, relevant sites and so on. Therefore, the purpose of this study is to find out a plan for an active use of it by Korean export and import firms and provide a number of policy suggestions that it can make best use of.

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Peroxynitrite Inactivates Carbonic Anhydrase II by Releasing Active Site Zinc Ion

  • Kim, Young-Mi;Han, Sang-Hwa
    • Bulletin of the Korean Chemical Society
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    • v.25 no.5
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    • pp.711-714
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    • 2004
  • Peroxynitrite enters erythrocytes through band 3 anion exchanger and oxidizes cytosolic proteins therein. As a protein associated with band 3, carbonic anhydrase II may suffer from peroxynitrite-induced oxidative damages. Esterase activity of carbonic anhydrase II decreased as the concentration of peroxynitrite increased. Neither hydrogen peroxide nor hypochlorite affected the enzyme activity. Inactivation of the enzyme was in parallel with the release of zinc ion, which is a component of the enzyme's active site. SDS-PAGE of peroxynitrite-treated samples showed no indication of fragmentation but non-denaturing PAGE exhibited new bands with lower positive charges. Western analysis demonstrated that nitration of tyrosine residues increased with the peroxynitrite concentration but the sites of nitration could not be determined. Instead MALDI-TOF analysis identified tryptophan-245 as a site of nitration. Such modification of tryptophan residues is responsible for the decrease in tryptophan fluorescence. These results demonstrate that peroxynitrite nitrates tyrosine and tryptophan residues of carbonic anhydrase II without causing fragmentation or dimerization. The peroxynitrite-induced inactivation of the enzyme is primarily due to the release of zinc ion in the enzyme's active site.

Self-organized Nanogels of Polysaccharide Derivatives in Anti-Cancer Drug Delivery

  • Park, Sin-Jung;Na, Kun
    • Journal of Pharmaceutical Investigation
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    • v.40 no.4
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    • pp.201-212
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    • 2010
  • Self-organized nanogels from polysaccharide derivatives offer a promising approach in treatment of cancer due to their flexibility in chemistry and their ability to improve the therapeutic index of a drug by modifying biodistribution by their preferential localization at target sites and lower distribution in normal healthy tissues. These properties have promoted studies of active cancer targeting by self-organized nanogels for even better accumulation in solid tumors. However although many researchers have reported their potential by using cell culture systems and small animal tumor models in cancer therapy, these nanogels need more decoration such as conjugation with targeting moiety and endowment of stimuli-sensitivity for precise targeting of the cancer site. In this review, we summarize the recent efforts in developing novel targeting approaches via active endocytosis and stimuli-sensitive systems responding to hyperthermic or acidic tumor pH conditions.

Highly Active Catalyst of Nickel Sulfate Supported on Titania for Ethylene Dimerization

  • Son, Jong Rak;Park, Won Cheon
    • Bulletin of the Korean Chemical Society
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    • v.22 no.12
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    • pp.1303-1308
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    • 2001
  • A series of catalysts, NiSO4/TiO2, for ethylene dimerization was prepared by the impregnation method using aqueous solution of nickel sulfate. On the basis of the results obtained from X-ray diffraction, the addition of NiSO4 shifted the transition of TiO2 from the anatase to the rutile phase toward higher temperatures due to the interaction between NiSO4 and TiO2. Nickel sulfate supported on titania was found to be very active even at room temperature. The high catalytic activity of NiSO4/TiO2 closely correlated with the increase of acidity and acid strength due to the addition of NiSO4. It is suggested that the active sites responsible for ethylene dimerization consist of low valent nickel, Ni+, with an acid.

Strain Development for the Over-production of Alkaline Protease from Vibrio metschnikovii by Molecular Evolution (분자진화 기술을 통한 Vibrio metschnikovii 유래 고활성 알칼리성 단백질 분해효소 생산균주 개발)

  • Shin, Yong-Uk;Lee, Gwa-Soo;Jo, Jae-Hyung;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.383-388
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    • 2010
  • Alkaline protease-overproducing strains of Vibrio metschnikovii were developed by using the molecular evolution from the classical mutants V. metschnikovii L12-23, N4-8, and KS1. Each vapK (Vibrio alkaline protease K) was obtained from the genomic DNAs of mutants by PCR to carry out the DNA shuffling. The modified vapK-1 obtained by DNA shuffling was used again as a template for the error-prone PCR to make the vapK-2. Both genes were cloned in the plasmid pKF3 to construct the recombinant plasmids which have one or two copies of the modified genes. The recombinant plasmids were back-transformed to V. metschnikovii KS1 to construct recombinant V. metschnikovii that expresses the alkaline protease. About 3.9-fold more protease activity was measured in the strain which has the plasmid containing two copies of vapK-2 when compared to strain KS1. When compared to wild type V. metschnikovii RH530, 43-fold more activity was achieved. Comparison of amino acids among vapK, vapK-1, and vapK-2 revealed that the active sites was highly conserved and not changed. However, many amino acids except the active sites were changed. These results suggested that the changes in amino acids might play an important role in the increase of protease activity by allowing the easy access of substrate to active sites of the protease. The fermentation of alkaline protease from the V. metschnikovii KS1 harboring the plasmid that contains two copies of vapK-1 showed the possibility of this strain to be used as industrial producer.

Relationship between noise and air pollution levels in Residential areas: comparison between roadside and non-roadside sites (소음과 대기오염도의 연계성에 대한 연구 - 일반주거지역의 도로 및 비도로지역의 비교)

  • Lee, Min-Hee;Park, Chan-Goo;Ryu, In-Cheol;Kim, Ki-Hyun
    • Analytical Science and Technology
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    • v.24 no.5
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    • pp.368-377
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    • 2011
  • In this study, we analyzed the relationship between noise and air pollution from 6 residential sites from 6 districts in Seoul. The survey of all noise date obtained from 6 sites were also compared between road and nonroad area and between active (06:00-22:00) and inactive time bands (22:00-06:00). Comparison of the data between two time bands indicates that their differences are not large enough. On the other hand, if comparison of data is made by correlation analysis for road and non-road areas, strong correlations between noise levels and air pollution tend to be prevalent consistently in road area relative to the non-road area. In contrast, a correlation between noise levels and air pollution tends to decrease at the road area in inactive period relative to active period.

The Composition and Emission Characteristics of VOCs from Major Waste Landfill Sites in Korea (전국 주요 쓰레기매립장 지역에 대한 VOC 성분의 조성과 배출특성에 대한 연구)

  • Kim, Ki-Hyun;Choi, Ye-Jin;SunWoo, Young
    • Journal of Environmental Policy
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    • v.4 no.1
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    • pp.69-91
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    • 2005
  • In order to accurately evaluate the distribution and emission characteristics of volatile organic compounds(VOCs) produced in the landfill environment, we conducted a series of field campaign to measure VOC emissions from a total of eight landfill sites located all across the Korean Peninsula. During the field measurements made from Sept. 2000 to Dec. 2004, we investigated VOC emission characteristics from 1 large scale, 5 mid scale, and 2 small scale landfill sites. The results from our 4-years study is used to provide valuable information for the management of landfill facilities in Korea. If annual emission rates of VOC are compared in terms of toluene, the magnitude of its emission differed between the closed and active landfills. Whereas closed landfills show emission rates from a few tens to 500g/yr, those of active ones were recorded as 1,000 to 40,000g/yr.

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