• Title/Summary/Keyword: biodesign

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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
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    • v.41 no.1
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    • pp.72-78
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    • 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.

Molecular Biodesign of Plant Leaves and Flowers

  • Kim Gyung-Tae
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.137-142
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    • 2003
  • The morphology of the leaves and the flowers of angiosperms exhibit remarkable diversity. One of the factors showing the greatest variability of leaf organs is the leaf index, namely, the ratio of leaf length to leaf width. In some cases, different varieties of a single species or closely related species can be distinguished by differences in leaf index. To some extent, the leaf index reflects the morphological adaptation of leaves to a particular environment. In addition, the growth of leaf organs is dependent on the extent of the expansion of leaf cells and on cell proliferation in the cellular level. The rates of the division and enlargement of leaf cells at each stage contribute to the final shape of the leaf, and play important roles throughout leaf development. Thus, the control of leaf shape is related to the control of the shape of cells and the size of cells within the leaf. The shape of flower also reflects the shape of leaf, since floral organs are thought to be a derivative of leaf organs. No good tools have been available for studies of the mechanisms that underlie such biodiversity. However, we have recently obtained some information about molecular mechanisms of leaf morphogenesis as a result of studies of leaves of the model plant, Arabidopsis thaliana. For example, the ANGUSTIFOLIA (AN) gene, a homolog of animal CtBP genes, controls leaf width. AN appears to regulate the polar elongation of leaf cells via control of the arrangement of cortical microtubules. By contrast, the ROTUNDIFOLIA3 (ROT3) gene controls leaf length via the biosynthesis of steroid(s). We provide here an overview of the biodiversity exhibited by the leaf index of angiosperms. Taken together, we can discuss on the possibility of the control of the shapes and size of plant organs by transgenic approaches with the results from basic researches. For example, transgenic plants that overexpressed a wildtype ROT3 gene had longer leaves than parent plants, without any changes in leaf width. Thus, The genes for leaf growth and development, such as ROT3 gene, should be useful tools for the biodesign of plant organs.

Molecular Biodesign of Plant Leaves and Flowers

  • Kim, Gyung-Tae
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2003.04a
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    • pp.49-55
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    • 2003
  • The morphology of the leaves and the flowers of angiosperms exhibit remarkable diversity. One of the factors showing the greatest variability of leaf organs is the leaf index, namely, the ratio of leaf length to leaf width. In some cases, different varieties of a single species or closely related species can be distinguished by differences in leaf index. To some extent, the leaf index reflects the morphological adaptation of leaves to a particular environment. In addition, the growth of leaf organs is dependent on the extent of the expansion of leaf cells and on cell proliferation in the cellular level. The rates of the division and enlargement of leaf cells at each stage contribute to the final shape of the leaf, and play important roles throughout leaf development. Thus, the control of leaf shape is related to the control of the shape of cells and the size of cells within the leaf. The shape of flower also reflects the shape of leaf, since floral organs are thought to be a derivative of leaf organs. No good tools have been available for studies of the mechanisms that underlie such biodiversity. However, we have recently obtained some information about molecular mechanisms of leaf morphogenesis as a result of studies of leaves of the model plant, Arabidopsis thaliana. For example, the ANGUSTIFOLIA (AN) gene, a homolog of animal CtBP genes, controls leaf width. AN appears to regulate the polar elongation of leaf cells via control of the arrangement of cortical microtubules. By contrast, the ROTUNDIFOLIA3 (ROT3) gene controls leaf length via the biosynthesis of steroid(s). We provide here an overview of the biodiversity exhibited by the leaf index of angiosperms. Taken together, we can discuss on the possibility of the control of the shapes and size of plant organs by transgenic approaches with the results from basic researches. For example, transgenic plants that overexpressed a wild-type ROT3 gene had longer leaves than parent plants, without any changes in leaf width. Thus, The genes for leaf growth and development, such as ROT3 gene, should be useful tools for the biodesign of plant organs.

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Immunosignature: Serum Antibody Profiling for Cancer Diagnostics

  • Chapoval, Andrei I;Legutki, J Bart;Stafford, Philip;Trebukhov, Andrey V;Johnston, Stephen A;Shoikhet, Yakov N;Lazarev, Alexander F
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.12
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    • pp.4833-4837
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    • 2015
  • Biomarkers for preclinical diagnosis of cancer are valuable tools for detection of malignant tumors at early stages in groups at risk and screening healthy people, as well as monitoring disease recurrence after treatment of cancer. However the complexity of the body's response to the pathological processes makes it virtually impossible to evaluate this response to the development of the disease using a single biomarker that is present in the serum at low concentrations. An alternative approach to standard biomarker analysis is called immunosignature. Instead of going after biomarkers themselves this approach rely on the analysis of the humoral immune response to molecular changes associated with the development of pathological processes. It is known that antibodies are produced in response to proteins expressed during cancer development. Accordingly, the changes in antibody repertoire associated with tumor growth can serve as biomarkers of cancer. Immunosignature is a highly sensitive method for antibody repertoire analysis utilizing high density peptide microarrays. In the present review we discuss modern methods for antibody detection, as well as describe the principles and applications of immunosignature in research and clinical practice.

A Series of IncQ-Based Reporter Plasmids for Use in a Range of Gram-Negative Genera

  • O'Sullivan, Laura E.;Nickerson, Cheryl A.;Wilson, James W.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.871-874
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    • 2010
  • Many studies require expression analysis of the same gene/promoter across a range of bacterial genera. However, there is currently a lack of availability of reporters based on the broad-host-range IncQ replicon, which is compatible with a popular improved IncP transfer system that is self-transfer defective. We report IncQ lacZ reporter plasmids with features including (1) compatibility with IncP, IncW, and pBHR/pBBR replicons, (2) a variety of antibiotic markers (Sp-r, Sm-r, Km-r, Cm-r), (3) convenient mobilization via a novel self-transfer-defective IncP conjugation system, and (4) GenBank DNA sequences. Utility is demonstrated using three different promoters in different Gram-negative genera.

Specific Binding and Catalytic Activation of the MAPK-MKP Complex

  • Kim, Myeongbin;Ryu, Seong Eon
    • Biodesign
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    • v.6 no.4
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    • pp.79-83
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    • 2018
  • Mitogen-activated protein kinases (MAPKs) are one of the most important enzymes in various cellular activities, and the MAPK signaling pathway is implicated in many disorders. MAPK phosphatases (MKPs) are regulators that contain a MAPK-binding domain (MBD) for MAPK recognition, and a catalytic domain (CD), for dephosphorylation and inactivation of MAPKs. Due to their crucial role in regulating the MAPK pathway, MKPs are regarded as a potential drug target in various diseases. Attempts have also been made to regulate the MAPK pathway by reducing the MKP activity. For drug development, it is important to understand the key features of MAPK-MKP complex formation. This review summarizes the studies on MAPK-MKP complexes, mainly focusing on their selective recognition and catalytic activation.

Crystallization and preliminary X-ray analysis of API5-FGF2 complex

  • Bong, Seoung Min;Lee, Byung Il
    • Biodesign
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    • v.6 no.4
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    • pp.92-95
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    • 2018
  • API5 is a unique oncogenic, non-BIR type IAP nuclear protein and is up-regulated in several cancers. It exerts several functions, such as apoptosis inhibition, cell cycle progression, cancer immune escape, and anticancer drug resistance. Although structural studies of API have revealed that API5 mediates protein-protein interactions, its detailed molecular functions remain unknown. Since FGF2 is one of API5's major interacting proteins, structural studies of the API5-FGF2 complex will provide insight into both proteins' molecular function. We overexpressed and purified API5 and FGF2 in Escherichia coli and crystallized the API-FGF2 complex using polyethylene glycol (PEG) 6000 as a precipitant. Diffraction data were collected to a $2.7{\AA}$ resolution using synchrotron X-rays. Preliminary diffraction analysis revealed that the API5-FGF2 complex crystal belongs to the space group $P2_12_12_1$ with the following unit cell parameters: a = 46.862, b = 76.523, $c=208.161{\AA}$. One asymmetric unit with 49.9% solvent contains one API5-FGF2 complex. Molecular replacement calculation, using API5 and FGF2 coordinates, provided a clear electron density map for an API5-FGF2 complex.

Purification, crystallization and X-ray crystallographic analysis of nicotinamidase Pnc1 from Kluyveromyces lactis

  • Kim, Shinae;Chang, Jeong Ho
    • Biodesign
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    • v.7 no.1
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    • pp.24-27
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    • 2019
  • Pnc1 converts nicotinamide to nicotinic acid to generate NAD+ through the Preiss-Handler pathway that is one of the NAD+-salvage pathway. By reducing levels of nicotinamide, an inhibitor of the NAD+-dependent histone deacetylase Sir2, yeast Pnc1 contributes gene silencing. In this study, to understand the structural features and molecular mechanism of nicotinamidase Pnc1, we overexpressed, purified, and crystallized the N-terminally His6-tagged Pnc1 protein from Kluyveromyces lactis and obtained X-ray diffraction data at a resolution of 2.2 Å. The crystals of the K. lactis Pnc1 (KlPnc1) belonged to space group P212121 with unit cell parameters a=38.5, b=77.3, c=83.3, and α=β=γ= 90°. There is one molecule in the asymmetric unit.

Kinetics of Photocatalytic Reactions with Porous Carriers Coated with Nano-$TiO_2$ Particles (나노-$TiO_2$ 입자로 코팅된 다공성 담체의 광촉매 반응에 관한 동력학)

  • Park, Seong-Jun;Rittmann, Bruce E.;Bae, Woo-Keun
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.10
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    • pp.927-932
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    • 2009
  • Toxic and recalcitrant organic pollutants in wastewaters can be effectively treated when advanced oxidation and biodegradation are combined, ideally with intimate coupling, in which both processes occur simultaneously in the same system. One means to achieve intimate coupling is to coat nanoscale $TiO_2$ on the outside of macroporous biofilm carriers. This study investigated the kinetics of photocatalysis with $TiO_2$-coated porous carriers. The carriers were made of polyvinyl alcohol (PVA) and coated with $TiO_2$ using a low-temperature sol-gel process. The $TiO_2$-coated carriers catalyzed the oxidation of methylene blue (MB) effectively under irradiation of UV light. The overall reaction rate with adsorption and photolysis saturated at high MB concentration, and approached the adsorption rate, which was first order for all MB concent rations. This result indicates that adsorbed MB may have slowed photocatalysis by blocking active sites for photocatalysis. The overall kinetics could be described by a quasi-Langmuir model. The estimated maximum specific (per unit mass of $TiO_2$) transformation rate of MB by the $TiO_2$-coated carriers was four times larger than that obtained from slurry-$TiO_2$ reactors. This observation demonstrated that the $TiO_2$ present as a coating on the carriers maintained high efficiency for transforming recalcitrant organic matter via photocatalysis. These findings serve as a foundation for advancement of an intimate coupling of photocatalysis to biodegradation.