• Title/Summary/Keyword: Genetic modification

Search Result 262, Processing Time 0.03 seconds

Oxidative modification of ferritin induced by hydrogen peroxide

  • Yoon, Jung-Hwan;An, Sung-Ho;Kyeong, Inn-Goo;Lee, Myeong-Seon;Kwon, Sang-Chul;Kang, Jung-Hoon
    • BMB Reports
    • /
    • v.44 no.3
    • /
    • pp.165-169
    • /
    • 2011
  • Excess free iron generates oxidative stress that may contribute to the pathogenesis of various causes of neurodegenerative diseases. In this study, we assessed the modification of ferritin induced by $H_2O_2$. When ferritin was incubated with $H_2O_2$, the degradation of ferritin L-chain increased with the $H_2O_2$ concentration whereas ferritin H-chain was remained. Free radical scavengers, azide, thiourea, and N-acetyl-$_L$-cysteine suppressed the $H_2O_2$-mediated ferritin modification. The iron specific chelator, deferoxamine, effectively prevented $H_2O_2$-mediated ferritin degradation in modified ferritin. The release of iron ions from ferritin was increased in $H_2O_2$ concentration-dependent manner. The present results suggest that free radicals may play a role in the modification and iron releasing of ferritin by $H_2O_2$. It is assumed that oxidative damage of ferritin by $H_2O_2$ may induce the increase of iron content in cells and subsequently lead to the deleterious condition.

Oxidative Modification of Neurofilament-L by Copper-catalyzed Reaction

  • Kim, Nam-Hoon;Kang, Jung-Hoon
    • BMB Reports
    • /
    • v.36 no.5
    • /
    • pp.488-492
    • /
    • 2003
  • Neurofilament-L (NF-L) is a major element of neuronal cytoskeletons and known to be important for neuronal survival in vivo. Since oxidative stress might play a critical role in the pathogenesis of neurodegenerative diseases, we investigated the role of copper and peroxide in the modification of NF-L. When disassembled NF-L was incubated with copper ion and hydrogen peroxide, then the aggregation of protein was proportional to copper and hydrogen peroxide concentrations. Dityrosine crosslink formation was obtained in copper-mediated NF-L aggregates. The copper-mediated modification of NF-L was significantly inhibited by thiol antioxidants, N-acetylcysteine, glutathione, and thiourea. A thioflavin-T binding assay was performed to determine whether the copper/$H_2O_2$ system-induced in vitro aggregation of NF-L displays amyloid-like characteristics. The aggregate of NF-L displayed thioflavin T reactivity, which was reminiscent of amyloid. This study suggests that copper-mediated NF-L modification might be closely related to oxidative reactions which may play a critical role in neurodegenerative diseases.

Oxidative modification of ferritin induced by methylglyoxal

  • An, Sung-Ho;Lee, Myeong-Seon;Kang, Jung-Hoon
    • BMB Reports
    • /
    • v.45 no.3
    • /
    • pp.147-152
    • /
    • 2012
  • Methylglyoxal (MG) was identified as an intermediate in non-enzymatic glycation and increased levels were reported in patients with diabetes. In this study, we evaluated the effects of MG on the modification of ferritin. When ferritin was incubated with MG, covalent crosslinking of the protein increased in a time- and MG dose-dependent manner. Reactive oxygen species (ROS) scavengers, $N-acetyl-_L-cysteine$ and thiourea suppressed the MG-mediated ferritin modification. The formation of dityrosine was observed in MG-mediated ferritin aggregates and ROS scavengers inhibited the formation of dityrosine. During the reaction between ferritin and MG, the generation of ROS was increased as a function of incubation time. These results suggest that ROS may play a role in the modification of ferritin by MG. The reaction between ferritin and MG led to the release of iron ions from the protein. Ferritin exposure to MG resulted in a loss of arginine, histidine and lysine residues. It was assumed that oxidative damage to ferritin caused by MG may induce an increase in the iron content in cells, which is deleterious to cells. This mechanism, in part, may provide an explanation or the deterioration of organs under diabetic conditions.

Examining the Gm18 and $m^1G$ Modification Positions in tRNA Sequences

  • Subramanian, Mayavan;Srinivasan, Thangavelu;Sudarsanam, Dorairaj
    • Genomics & Informatics
    • /
    • v.12 no.2
    • /
    • pp.71-75
    • /
    • 2014
  • The tRNA structure contains conserved modifications that are responsible for its stability and are involved in the initiation and accuracy of the translation process. tRNA modification enzymes are prevalent in bacteria, archaea, and eukaryotes. tRNA Gm18 methyltransferase (TrmH) and tRNA $m^1G37$ methyltransferase (TrmD) are prevalent and essential enzymes in bacterial populations. TrmH involves itself in methylation process at the 2'-OH group of ribose at the 18th position of guanosine (G) in tRNAs. TrmD methylates the G residue next to the anticodon in selected tRNA subsets. Initially, $m^1G37$ modification was reported to take place on three conserved tRNA subsets ($tRNA^{Arg}$, $tRNA^{Leu}$, $tRNA^{Pro}$); later on, few archaea and eukaryotes organisms revealed that other tRNAs also have the $m^1G37$ modification. The present study reveals Gm18, $m^1G37$ modification, and positions of $m^1G$ that take place next to the anticodon in tRNA sequences. We selected extremophile organisms and attempted to retrieve the $m^1G$ and Gm18 modification bases in tRNA sequences. Results showed that the Gm18 modification G residue occurs in all tRNA subsets except three tRNAs ($tRNA^{Met}$, $tRNA^{Pro}$, $tRNA^{Val}$). Whereas the $m^1G37$ modification base G is formed only on $tRNA^{Arg}$, $tRNA^{Leu}$, $tRNA^{Pro}$, and $tRNA^{His}$, the rest of the tRNAs contain adenine (A) next to the anticodon. Thus, we hypothesize that Gm18 modification and $m^1G$ modification occur irrespective of a G residue in tRNAs.

Structural Dynamic Optimization Using a Genetic Algorithm(GA) (유전자 알고리즘(GA)을 이용한 구조물의 동적해석 및 최적화)

  • 이영우;성활경
    • Journal of the Korean Society for Precision Engineering
    • /
    • v.17 no.5
    • /
    • pp.93-99
    • /
    • 2000
  • In many dynamic structural optimization problems, the goal is to reduce the total weight of the structure without causing the resonance. Up to now, gradient informations(i.e., design sensitivity) have been used to achieve the goal. For some class of dynamic problems, especially coalescent eigenvalue Problems with multiobjective optimization, the design sensitivity analysis is too much complicated mathematically and numerically. Therefore, this article proposes a new technique fur structural dynamic modification using a mode modification method with Genetic Algorithm(GA). In GA formulation, fitness is defined based on penalty function approach. Design variables are iteratively improved by using genetic algorithm. Two numerical examples are shown, (ⅰ) a cantilevered plate, and (ⅱ) H-shaped structure. The results demonstrate that the proposed method is highly efficient.

  • PDF

Protective Effects of Histidine Dipeptides on the Modification of Neurofilament-L by the Cytochrome c/Hydrogen Peroxide System

  • Kim, Nam-Hoon;Kang, Jung-Hoon
    • BMB Reports
    • /
    • v.40 no.1
    • /
    • pp.125-129
    • /
    • 2007
  • Neurofilament-L (NF-L) is a major element of the neuronal cytoskeleton and is essential for neuronal survival. Moreover, abnormalities in NF-L result in neurodegenerative disorders. Carnosine and the related endogeneous histidine dipeptides prevent protein modifications such as oxidation and glycation. In the present study, we investigated whether histidine dipeptides, carnosine, homocarnosine, or anserine protect NF-L against oxidative modification during reaction between cytochrome c and $H_2O_2$. Carnosine, homocarnosine and anserine all prevented cytochrome c/$H_2O_2$-mediated NF-L aggregation. In addition, these compounds also effectively inhibited the formation of dityrosine, and this inhibition was found to be associated with the reduced formations of oxidatively modified proteins. Our results suggest that carnosine and histidine dipeptides have antioxidant effects on brain proteins under pathophysiological conditions leading to degenerative damage, such as, those caused by neurodegenerative disorders.

The Roles of Tryptophan and Histidine Residues in the Catalytic Activities $\beta$-Cyclodextrin Glucanotransferase from Bacillus firmus var. alkalophilus

  • Shin, Hyun-Dong;Kim, Chan;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
    • /
    • v.9 no.1
    • /
    • pp.62-69
    • /
    • 1999
  • In order to investigate the critical amino acid residues involved in the catalytic activities of $\beta$-cyclodextrin glucanotransferase ($\beta$-CGTase) excreted by Bacillus firmus var. alkalophilus, the amino acid residues in $\beta$-CGTase were modified by various site-specific amino acid modifying reagents. The cyclizing and amylolytic activities of $\beta$-CGTase were all seriously reduced after treatment with Woodward's reagent K (WRK) modifying aspartic/glutamic acid, N-bromosuccinimde (NBS) modifying tryptophan, and diethylpyrocarbonate (DEPC) modifying histidine residues. The roles of tryptophan and histidine residues in $\beta$-CGTase were further investigated by measuring the protection effect of various substrates during chemical modification, comparing protein mobility in native and affinity polyacrylamide gel electrophoresis containing soluble starch, and comparing the $K_m$ and $V_{max}$ values of native and modified enzymes. Tryptophan residues were identified as affecting substrate-binding ability rather than influencing catalytic activities. On the other hand, histidine residues influenced catalytic ability rather than substrate-binding ability, plus histidine modification had an effect on shifting the optimum pH and pH stability.

  • PDF

Tree Biotechnology and Environmental Concerns

  • Kant, Tarun;Emmanuel, C.J.S.K.
    • Journal of Plant Biotechnology
    • /
    • v.6 no.1
    • /
    • pp.1-7
    • /
    • 2004
  • Forestry sector has witnessed some unprecedented events in the recent past both in terms of galloping biotechnological developments and heated environmental debates over risks associated with release of transgenic trees. Improvements in the in vitro propagation techniques has made it possible to develop tissue culture based plant regeneration protocols just for about any tree species. And with the inclusion of every new species within the realms of tissue culture technology, it becomes a candidate for genetic improvement through recombinant DNA technology, the so called genetic engineering. Poplars and their hybrids serve as the model tree species on which most of the genetic transformation work as been carried out. A lot of work has also gone in genetic transformation of fruit trees and trees of horticultural interests. Trees have been successfully transformed for traits ranging from reduction of length of juvenile phase to alteration of tree architecture to altering wood quality by lignin and cellulose modification. More-over trees have been genetically engineered successfully to combat various types of insect pests and pathogens causing diseases. But all these developments have ignited controversies over the possible benefits and risks associated with transgenic plantations by various environmental agencies and activists. Solutions to most of these concerns can be found out with more intensive prioritized research.

Structural Dynamic Optimization of Diesel Generator systems Using Genetic Algorithm(GA) (유전자 알고리즘을 이용한 선박용 디젤발전기 시스템의 동특성 해석 및 최적화)

  • 이영우;성활경
    • Journal of Advanced Marine Engineering and Technology
    • /
    • v.24 no.3
    • /
    • pp.99-105
    • /
    • 2000
  • For multi-body dynamic problems. especially coalescent eigenvalue problems with multiobjective optimization, the design sensitivity analysis is too much complicated mathematically and numerically. Therefore, this article proposes a new technique for structural dynamic modification using a mode modification and homologous structures design method with Genetic Algorithm(GA). In this work, the homologous structure of the resiliently mounted multi-body for marine diesel generator systems is studied and the problem is treated as a combinational optimization problem using the GA. In GA formulation, fitness is defined based on penalty function approach. That include homology, allowable stress and minimum weight of common plate.

  • PDF