Synthesis and biological activities of 2-functionalized purine nucleosides

Novel purine nucleosides functionalized at the 2-position have been prepared using new applications of synthetic methodology. The target molecules were designed as potential inhibitors (as their monophosphates) of the enzyme, inosine monophosphate dehydrogenase (IMPDH), and representative inhibition data are presented. Antiviral data of the compounds are discussed.     

SCAR Marker Development for the Identification of Elite Germplasm of Moringa Oleifera Lam.-A Never Die Plant

Plant Molecular Biology Reporter - Moringa oleifera Lam. (drumstick) belongs to the family Moringaceae that is originated from sub-Himalayan tracts of Northern India distributed worldwide in the...     

Crystal Structure of Diaminopimelate Epimerase from Arabidopsis thaliana, an Amino Acid Racemase Critical for l-Lysine Biosynthesis

Diaminopimelate (DAP) epimerase is a key enzyme for the biosynthesis of lysine in plants. Lysine is an essential dietary nutrient for mammals. In both plants and bacteria, DAP epimerase catalyzes the interconversion of ll-DAP and dl(meso)-DAP. The absence of a mammalian homolog makes DAP epimerase a promising target for the design of novel herbicides and antibacterials. This enzyme requires no cofactors and it functions through an unusual mechanism involving two cysteine residues acting in concert and alternating as a base (thiolate) and as an acid (thiol). The present study reports the crystal structures of two enzyme-inhibitor complexes of DAP epimerase from Arabidopsis thaliana with different isomers of the irreversible inhibitor and substrate mimic, 2-(4-amino-4-carboxybutyl)-aziridine-2-carboxylate, at 1.95 and 2.3 Å resolution. These structures provide the first atomic details of a plant amino acid racemase. Structural analysis reveals that ligand binding to a cleft between the two domains of the enzyme is accompanied by domain closure with two strictly conserved cysteine residues, Cys99 and Cys254, optimally positioned to perform acid/base catalysis via a carbanion stabilization mechanism on the stereogenic α-carbon atom of the amino acid. Stereochemical control in catalysis is achieved by means of a highly symmetric catalytic site that can accommodate both the l and d stereogenic centers of DAP at the proximal site, whereas specific interactions at the distal site require only the l configuration. Structural comparisons of the plant enzyme with its bacterial counterpart from Haemophilus influenzae reveal significant conservation of amino acid residues around the active site that extends to their three-dimensional structures and catalytic mechanism.     

A role of unliganded thyroid hormone receptor in postembryonic development in Xenopus laevis

A fascinating feature of thyroid hormone (T3) receptors (TR) is that they constitutively bind to promoter regions of T3-response genes, providing dual functions. In the presence of T3, TR activates T3-inducible genes, while unliganded TR represses these same genes. Although this dual function model is well demonstrated at the molecular level, few studies have addressed the presence or the role of unliganded TR-induced repression in physiological settings. Here, we analyze the role of unliganded TR in Xenopus laevis development. The total dependence of amphibian metamorphosis upon T3 provides us a valuable opportunity for studying TR function in vivo. First, we designed a dominant negative form of TR-binding corepressor N-CoR (dnN-CoR) consisting of its receptor interacting domain. We confirmed its dominant negative activity by showing that dnN-CoR competes away the binding of endogenous N-CoR to unliganded TR and relieves unliganded TR-induced gene repression in frog oocytes. Next, we overexpressed dnN-CoR in tadpoles through transgenesis and analyzed its effect on gene expression and development. Quantitative RT-PCR revealed significant derepression of T3-response genes in transgenic animals. In addition, transgenic tadpoles developed faster than wild type siblings, with an acceleration of as much as 7 days out of the 30-day experiment. These data thus provide in vivo evidence for the presence and a role of unliganded TR-induced gene repression in physiological settings and strongly support our earlier model that unliganded TR represses T3-response genes in premetamorphic tadpoles to regulate the progress of development.     

The immune landscape in tuberculosis reveals populations linked to disease and latency

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Isolation of 4-hydroxy 3-methyl 2-butenyl 4-diphosphate reductase (ApHDR) gene of methyl erythritol diphosphate (MEP) pathway,in silico analysis and differential tissue specific ApHDR expression in Andrographis paniculata (Burm. f) Nees

The full length Andrographis paniculate 4-hydroxy 3-methyl 2-butenyl 4-diphosphate reductase (ApHDR) gene of MEP pathway was isolated for the first time. The ApHDR ORF with 1404 bp flanked by 100 bp 5′UTR and 235 bp 3′UTR encoding 467 amino acids (NCBI accession number: {"type":"entrez-nucleotide","attrs":{"text":"MK503970","term_id":"1732262198"}}MK503970) and cloned in pET 102, transformed and expressed in E. coli BL21. The ApHDR protein physico-chemical properties, secondary and tertiary structure were analyzed. The Ramachandran plot showed 93.8% amino acids in the allowed regions, suggesting high reliability. The cluster of 16 ligands for binding site in ApHDR involved six amino acid residues having 5–8 ligands. The Fe-S cluster binding site was formed with three conserved residues of cysteine at positions C123, C214, C251 of ApHDR. The substrate HMBPP and inhibitors clomazone, paraquat, benzyl viologen’s interactions with ApHDR were also assessed using docking. The affinity of Fe-S cluster binding to the cleft was found similar to HMBPP. The HPLC analysis of different type of tissue (plant parts) revealed highest andrographolide content in young leaves followed by mature leaves, stems and roots. The differential expression profile of ApHDR suggested a significant variation in the expression pattern among different tissues such as mature leaves, young leaves, stem and roots. A 16-fold higher expression of ApHDR was observed in the mature leaves of A. paniculata as compared to roots. The young leaves and stem showed 5.5 fold and fourfold higher expression than roots of A. paniculata. Our result generated new genomic information on ApHDR which may open up prospects of manipulation for enhanced diterpene lactone andrographolide production in A. paniculata.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-00952-0.     

A comparative study on permeabilization treatments for in situ determination of phytase of Rhodotorula gracilis

Rhodotorula gracilis , a red yeast, was shown to produce phytase and the cultural conditions were standardized for phytase production. Permeabilization treatments using detergents, organic solvents, enzymes and physical methods were performed and compared by measuring the phytase activity of whole cells. None of the permeabilization methods except the freeze-thaw method enhanced phytase activity. The enzyme activity of a yeast suspension subjected to 15 cycles of freezing and thawing was 77·26 U g⁻¹ cells. Phytate in feeds is an anti-nutritional factor which complexes with many minerals, rendering it unavailable to monogastric animals. Rhodotorula gracilis subjected to freeze-thawing can be used as a source of phytase in feeds to enhance phosphorus availability.     

Role of aminolevulinic acid in improving biomass production in Vigna catjung, V. mungo, and V. radiata

A precursor in tetrapyrrole biosynthesis, 5-aminolevulinic acid (ALA), was applied via presowing soaking in Vigna catjung, V. mungo, and V. radiata. ALA increased plant growth and influenced dry matter accumulation in leaves, stems, and pods through increased chlorophyll content and photosynthetic CO₂ absorption. At harvest, ALA treated plants had increased number of pods per plant, seeds per pod, 100 seed dry matter, biological yield, and the harvest index. Therefore, pretreatment of seeds with optimal concentration of ALA is recommended for improving the growth and productivity of tropical legumes. This revised version was published online in July 2006 with corrections to the Cover Date.