• Title/Summary/Keyword: Synthetic biology

Search Result 371, Processing Time 0.023 seconds

Effect of Ganglioside $G_{M3}$ on the Erythrocyte Glucose Transporter (GLUT1): Conformational Changes Measured by Steady-State and Time-Resolved Fluorescence Spectroscopy

  • Yoon, Hae-Jung;Lee, Min-Yung;Jhon, GiI-Ja
    • BMB Reports
    • /
    • v.30 no.4
    • /
    • pp.240-245
    • /
    • 1997
  • Interactions between ganglioside $G_{M3}$ and glucose transporter, GLUT1 were studied by measuring the effect of $G_{M3}$ on steady-state and time-resolved fluorescence of purified GLUT1 in synthetic lipids and on the 3-O-methylglucose uptake by human erythrocytes. The intrinsic tryptophan fluorescence showed a GLUT 1 emission maximum of 335 nm, and increased in the presence of $G_{M3}$ by 12% without shifting the emission maximum, The fluorescence lifetimes of intrinsic tryptophan on GLUT1 consisted of a long component of 7.8 ns and a short component of 2,3 ns and $G_{M3}$ increased both lifetime components. Lifetime components were quenched by acrylamide and KI. Acrylarnide-mduced quenching of long-lifetime components was partly recovered by $G_{M3}$ However. KI-induccd quenching of short- and long-lifetime components was not rescued by $G_{M3}$. The anisotropy of 1.6-diphenyl-1.3.5-hexatriene (DPH)-probed dimyristoylphosphatidylcholine (DMPC) model membrane was also increased with $G_{M3}$ incorporation, The transport rate of 3-O-methylglucose increased by 20% with $G_{M3}$ incorporation on the erythrocytes, Therefore, $G_{M3}$ altered the environment of lipid membrane and induced the conformational change of GLUT1.

  • PDF

Replacement of the antifreeze-like domain of human N-acetylneuraminic acid phosphate synthase with the mouse antifreeze-like domain impacts both N-acetylneuraminic acid 9-phosphate synthase and 2-keto-3-deoxy-D-glycero-Dgalacto- nonulosonic acid 9-phosphate synthase activities

  • Reaves, Marshall Louis;Lopez, Linda Carolyn;Daskalova, Sasha Milcheva
    • BMB Reports
    • /
    • v.41 no.1
    • /
    • pp.72-78
    • /
    • 2008
  • Human NeuNAc-9-P synthase is a two-domain protein with ability to synthesize both NeuNAc-9-P and KDN-9-P. Its mouse counterpart differs by only 20 out of 359 amino acids but does not produce KDN-9-P. By replacing the AFL domain of the human NeuNAc-9-P synthase which accommodates 12 of these differences, with the mouse AFL domain we examined its importance for the secondary KDN-9-P synthetic activity. The chimeric protein retained almost half of the ability of the human enzyme for KDN-9-P synthesis while the NeuNAc-9-P production was reduced to less than 10%. Data from the homology modeling and the effect of divalent ions and temperature on the enzyme activities suggest conformational differences between the human and mouse AFL domains that alter the shape of the cavity accommodating the substrates. Therefore, although the AFL domain itself does not define the ability of the human enzyme for KDN-9-P synthesis, it is important for both activities by aiding optimal positioning of the substrates.

Identification of a lead small-molecule inhibitor of anthrax lethal toxin by using fluorescence-based high-throughput screening

  • Wei, Dong;Bu, Zhaoyun;Yu, Ailian;Li, Feng
    • BMB Reports
    • /
    • v.44 no.12
    • /
    • pp.811-815
    • /
    • 2011
  • Inhalational anthrax is caused by B. anthracis, a virulent sporeforming bacterium which secretes anthrax toxins consisting of protective antigen (PA), lethal factor (LF) and edema factor (EF). LF is a Zn-dependent metalloprotease and is the main determinant in the pathogenesis of anthrax. Here we report the identification of a lead small-molecule inhibitor of anthrax lethal factor by screening an available synthetic small-molecule inhibitor library using fluorescence-based high-throughput screening (HTS) approach. Seven small molecules were found to have inhibitory effect against LF activity, among which SM157 had the highest inhibitory activity. All theses small molecule inhibitors inhibited LF in a noncompetitive inhibition mode. SM157 and SM167 are from the same family, both having an identical group complex, which is predicted to insert into S1' pocket of LF. More potent small-molecule inhibitors could be developed by modifying SM157 based on this identical group complex.

Change in Photosynthesis, Proline Content, and Osmotic Potential of Corn Seedling under High-Saline Condition

  • Yoon Byeong Sung;Jin Chengwn;Park Sang Un;Cho Dong Ha
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.50 no.1
    • /
    • pp.28-31
    • /
    • 2005
  • To identify salt-tolerance characteristics of corn seedling was treated in solution of 0, 50 and 100 mM NaCl of hydroponic cultivation. In photosynthesis of corn seedling, there was no large difference between 50mM and 0 mM NaCl solution, however, in 100 mM NaCl solution, the tolerance gradually decreased to $76\%,\;49\%,\;and\;31\%$ after one day, four days, and seven days, respectively, in comparison to 0 mM NaCl solution. Osmotic potential of corn in seedling period was significantly decreased with increasing saline level, however, free proline content in the plant on the ground was significantly increased with increasing saline level and with the lapse of time. In terms of correlation among major characteristics, there was a highly significant positive difference between osmotic pressure potential and photosynthesis, However, highly negative correlation was found between osmotic pressure potential and free proline content. In addition, it was expected that young seedling of corn with saline tolerance may be utilized in the transplantation in salt-accumulated land. Based on above-shown result, in terms of saline tolerance of Chalok-2 variety, growth suppression was serious with 100mM NaCl solution. However, growth was expected that seedling growth would be favorable under 50 mM NaCl solution.

Protein Synthesis Pattern Analysis in the Regenerating Salamander Limb

  • Ju, Bong-Gun;Kim, Won-Sun
    • Animal cells and systems
    • /
    • v.4 no.2
    • /
    • pp.145-150
    • /
    • 2000
  • Retinoic acid (RA) evokes pattern duplication in the regenerating salamander limb. Interestingly, it also enhances dedifferentiation in the regenerate by the morphological, histological and biochemical criteria. To examine whether there is any correlation between the RA-evoked pattern duplication and de novo protein synthetic profile in the regenerating salamander limb, especially during dedifferentiation, we analyzed stage-specific protein synthesis pattern in the normal and RA-treated regenerating limbs by metabolic labeling followed by two-dimensional gel electrophoresis. In the regenerating limbs without RA treatment, a few hundred kinds of proteins were found to be synthesized at the stage of wound healing and the total number of protein synthesized increased greatly as regeneration proceeded. The same trend was also observed in the RA-treated regenerating limbs. Interestingly, some protein spots were noted to be either newly synthesized or highly expressed by the RA treatment especially at the stage of dedifferentiation. The results shows that the enhancement of dedifferentiation state after the RA treatment correlates well with the protein synthesis profile, and suggest that those proteins are important for the RA-evoked pattern duplication in the regenerating limbs of salamander.

  • PDF

Cytotoxic and Antioxidant Effects of Taraxacum coreanum Nakai. and T. officinale WEB. Extracts (흰민들레와 서양민들레 추출물의 세포독성 및 항산화 활성 비교)

  • Lee, Hyun-Hwa;Lee, Sook-Yeong
    • Korean Journal of Medicinal Crop Science
    • /
    • v.16 no.2
    • /
    • pp.79-85
    • /
    • 2008
  • Some studies of dandelion have been carried out on bioactivities, however, no comparative analysis on antioxidant and cytotoxic activities in the Korean dandelion (T. coreanum NAKAI.; KD) and dandelion (T. officinale WEB.) have been reported. In this study, the extracts of KD and dandelion analyzed relative phenolic contents and free radical scavenging, antioxidant enzyme and cytotoxic activities. The extract of Korean dandelion leaves (KDL) exhibited a higher phenolics content ($368{\pm}11.5\;mg/l00\;g$) and a strong free radical scavenging activity ($RC_{50}$value;87.89 ug/ml) than other parts and BHT (120.12 ug/ml), synthetic antioxidant. At ascorbate peroxidase (APX) activity, the dandelion root (DR) had a greater (38.8 U/mg protein) antioxidant enzyme compare to the dandelion leaves (12.2 U/mg protein). The catalase (CAT) and superoxide dismutase (SOD) followed higher enzyme activity in Korean dandelion root (KDR) than other parts. In a cytotoxic activity against human cancer cell, the extracts of KDR was found to be active against Calu-6, HCT-116, and SNU-601 cell lines, with $IC_{50}$ values of 522.34, 532.74 and 614.85 ug/ml, respectively. These results suggest that KD and dandelion would be an alternative antioxidant source, based on natural plant resources.

The Synthesis of $\alpha$-lhnctionalized Benzykmine ($\alpha$-Functionalized Benzylamine의 합성)

  • Kim, Hyeung-Jae;Cho, Su-Dong;Ahn, Chul Jin;Joo, Woo-Hong;Shin, Dong Soo
    • Journal of the Korean Chemical Society
    • /
    • v.44 no.5
    • /
    • pp.442-447
    • /
    • 2000
  • $\alpha$-Functionalized benzylamine(Nu=OAc, OMe, N$_3$) will be very usefud as a starting material or synton in organic synthesis. We have investigated the first synthetic methodology of phthalimide-ben-zylacetate from benzylphthalimide, which are prepared in NBS/NaOAc/HOAc in chlorobenzene at 137$^{\circ}C$ for 12 hr in good yield. Finally, amino(phenyl)methyl acetate (1) was synthesized from phthalimide-benzyl acetate under NH$_2$NH$_2$/HOAC.

  • PDF

Mechanisms of herbicide resistance in weeds

  • Bo, Aung Bo;Won, Ok Jae;Sin, Hun Tak;Lee, Jeung Joo;Park, Kee Woong
    • Korean Journal of Agricultural Science
    • /
    • v.44 no.1
    • /
    • pp.1-15
    • /
    • 2017
  • In major field crops, synthetic herbicides have been used to control weeds worldwide. Globally, herbicide resistance in weeds should be minimized because it is a major limiting factor for food security. Cross resistance can occur with herbicides within the same or in different herbicide families and with the same or different sites of action. Multiple resistance refers to evolved mechanisms of resistance to more than one herbicide (e.g., resistance to both ALS-inhibitors and ACCase-inhibitors) and this resistance was brought about by separate selection processes. Target site resistance could occur from changes at the biochemical site of action of one herbicide. Non target site resistance occurs through mechanisms which reduce the number of herbicide molecules that reach the herbicide target site. There are currently 480 unique cases (species ${\times}$ site of action) of herbicide resistance globally in 252 plant species (145 dicots and 105 monocots). To date, resistance in weeds has been reported to 161 different herbicides, involving 23 of the 26 known herbicide sites of action. Finally, it can be concluded that we can protect crops associated to herbicide resistant weeds by applications of biochemical, genetic and crop control strategies.

Induction of Peripheral Tolerance in Dual TCR T Cells: an Evidence for Non-dominant Signaling by One TCR

  • Hah, Chae-Rim;Kim, Mi-Hyung;Kim, Kil-Hyoun
    • BMB Reports
    • /
    • v.38 no.3
    • /
    • pp.334-342
    • /
    • 2005
  • Recently, the existence of T cells with dual T cell receptor (TCR) in the immune system is generally accepted, while it has been controversial whether signals through one TCR would affect the functions of the other. In this study T cells expressing two different TCR were obtained from cross-hybrids of LCMV and AND TCR transgenic mice specific for the gp33 and peptide fragment of PCC (fPCC), respectively. Peptide stimulation demonstrated that the dual TCR T cells functioned independently in an antigen-specific manner. To examine whether the tolerance targeted for the one TCR affects the responsiveness of the other, the cross-hybrids were treated with gp33. Although T cells from F1 mice were rendered anergenic to gp33, no functional changes to fPCC were observed in terms of cellular proliferation and IL-2 secretion, suggesting that the dual TCR T cells remained reactive to fPCC. We therefore propose that signaling through the TCR is receptor-specific and 'negative dominance' of one TCR by tolerance induction is not applicable in this dual TCR system.

Antibacterial Activities of Peptides Designed as Hybrids of Antimicrobial Peptides

  • Shin, Song-Yub;Kang, Joo-Hyun;Lee, Myung-Kyu;Hahm, Kyung-Soo
    • BMB Reports
    • /
    • v.29 no.6
    • /
    • pp.545-548
    • /
    • 1996
  • CA(1-8)ME(1-12), the CA-ME hybrid peptide of the amino terminal segments of cecropin A (CA) and melittin (ME), has been reported to have a broad spectrum and improved potency without a hemolytic property. In order to obtain new synthetic peptides with powerful antibacterial activity without hemolytic activity, several hybrid peptides were designed from the sequences of CA, ME, magainin 2, bombinin and lactoferricin. All hybrid peptides were constructed to form an amphipathically basic-flexible-hydrophobic structure and synthesized by the solid phase method. Their hemolytic activities against human red blood cells and antibacterial activities against both Gram-positive and Gram-negative bacteria were detennined. CA(1-8)MA(1-12), CA(1-8)BO(1-12), MA(10-17)ME(1-12) and LF(20-29)ME(1-12) showed comparable activities with broad spectra against both Gram-positive and Gram-negative bacteria relative to CA(1-8)ME(1-12) but without hemolytic properties. These hybrid peptides, therefore, could be useful as model peptides to design a novel peptide with improved antibacterial activity and study on structure-activity relationships of antimicrobial peptides.

  • PDF