Search results for "AMINO ACID"
showing 10 items of 3965 documents
Experimental Evolution Reveals a Genetic Basis for Membrane-Associated Virus Release
2021
Many animal viruses replicate and are released from cells in close association to membranes. However, whether this is a passive process or is controlled by the virus remains poorly understood. Importantly, the genetic basis and evolvability of membrane-associated viral shedding have not been investigated. To address this, we performed a directed evolution experiment using coxsackievirus B3, a model enterovirus, in which we repeatedly selected the free-virion or the fast-sedimenting membrane-associated viral subpopulations. The virus responded to this selection regime by reproducibly fixing a series of mutations that altered the extent of membrane-associated viral shedding, as revealed by fu…
Main protein component of high density lipoprotein, apolipoprotein A1, modulates leukocyte cell respiration
2015
HDL:n (high density lipoprotein) pääproteiinikomponentin, apolipoproteiini A1:n (ApoA1), on todettu lisäävän glykolyysia ja oksidatiivista fosforylaatiota hiiren luurankolihassoluissa. ApoA1:llä oletetaan myös olevan merkittävä rooli lihaksen normaalissa aineenvaihdunnassa ja siihen liittyvissä sairauksissa. ApoA1 vaikuttaa leukosyyttien aktivaatioon ja jakautumiseen säätelemällä solun sisäistä kolesterolitasoa. Nämä muutokset vaikuttavat myös solun aineenvaihduntaan. ApoA1:n suoranaista vaikutusta leukosyyttien soluhengitykseen ei kuitenkaan ole aiemmin tutkittu. Aiemmin on esitetty, että leukosyyttejä voitaisiin käyttää biomarkkerina kuvaamaan muutoksia mitokondrioiden toiminnassa aineenv…
The role of lipid metabolism in the acquisition of desiccation tolerance inCraterostigma plantagineum: a comparative approach
2013
Summary Dehydration leads to different physiological and biochemical responses in plants. We analysed the lipid composition and the expression of genes involved in lipid biosynthesis in the desiccation-tolerant plant Craterostigma plantagineum. A comparative approach was carried out with Lindernia brevidens (desiccation tolerant) and two desiccation-sensitive species, Lindernia subracemosa and Arabidopsis thaliana. In C. plantagineum the total lipid content remained constant while the lipid composition underwent major changes during desiccation. The most prominent change was the removal of monogalactosyldiacylglycerol (MGDG) from the thylakoids. Analysis of molecular species composition rev…
Factors affecting the production of putrescine from agmatine by Lactobacillus hilgardii X1B isolated from wine
2008
Aims: To elucidate and characterize the metabolic putrescine synthesis pathway from agmatine by Lactobacillus hilgardii X1B. Methods and Results: The putrescine formation from agmatine by resting cells (the normal physiological state in wine) of lactic acid bacteria isolated from wine has been determined for the first time. Agmatine deiminase and N-carbamoylputrescine hydrolase enzymes, determined by HPLC and LC-Ion Trap Mass Spectrometry, carried out the putrescine synthesis from agmatine. The influence of pH, temperature, organic acids, amino acids, sugars and ethanol on the putrescine formation in wine was determined. Conclusions: Resting cells of Lact. hilgardii X 1B produce putrescine …
Kinetic evidence for the incorporation of the [(pentamethylcyclopentadienyl) (2,2′-bipyridyl)(aquo)rhodium(III)] complex into DPPC vesicles
2008
Abstract The binding of the [(pentamethylcyclopentadienyl) (2,2′-bipyridyl)(aquo)rhodium(III)] complex [Cp*RhIII(bpy)H2O]2+, to l -α-dipalmitoylphosphatidyl choline (DPPC) vesicles has been estimated by studying the kinetics of the electron transfer reaction between the rhodium(III) complex and formiate ions. Kinetic measurements carried out under anaerobic conditions in absence and presence of DPPC show that the total reaction is composed of two steps. The rate of the first reaction increases with the phospholipid concentration, while that of the second process is independent of the concentration of DPPC. This is consistent with a reaction, where the two reacting species are partitioned be…
Single-chain antibodies against a plant viral RNA-dependent RNA polymerase confer virus resistance.
2004
Crop loss due to viral diseases is still a major problem for agriculture today. We present a strategy to achieve virus resistance based on the expression of single-chain Fv fragments (scFvs) against a conserved domain in a plant viral RNA-dependent RNA polymerase (RdRp), a key enzyme in virus replication. The selected scFvs inhibited complementary RNA synthesis of different plant virus RdRps in vitro and virus replication in planta. Moreover, the scFvs also bound to the RdRp of the distantly related hepatitis C virus. T(1) and T(2) progeny of transgenic lines of Nicotiana benthamiana expressing different scFvs either in the cytosol or in the endoplasmic reticulum showed varying degrees of r…
Modulation of Hepatitis C Virus NS5A Hyperphosphorylation by Nonstructural Proteins NS3, NS4A, and NS4B
1999
NS5A of the hepatitis C virus (HCV) is a highly phosphorylated protein involved in resistance against interferon and required most likely for replication of the viral genome. Phosphorylation of this protein is mediated by a cellular kinase(s) generating multiple proteins with different electrophoretic mobilities. In the case of the genotype 1b isolate HCV-J, in addition to the basal phosphorylated NS5A (designated pp56), a hyperphosphorylated form (pp58) was found on coexpression of NS4A (T. Kaneko, Y. Tanji, S. Satoh, M. Hijikata, S. Asabe, K. Kimura, and K. Shimotohno, Biochem. Biophys. Res. Commun. 205:320‐326, 1994). Using a comparative analysis of two full-length genomes of genotype 1b…
Functional properties of a monoclonal antibody inhibiting the hepatitis C virus RNA-dependent RNA polymerase.
2001
The hepatitis C virus (HCV) RNA-dependent RNA polymerase (RdRp), represented by nonstructural protein 5B (NS5B), has recently emerged as a promising target for antiviral intervention. Here, we describe the isolation, functional characterization, and molecular cloning of a monoclonal antibody (mAb) inhibiting the HCV RdRp. This mAb, designated 5B-12B7, binds with high affinity to a conformational epitope in the palm subdomain of the HCV RdRp and recognizes native NS5B expressed in the context of the entire HCV polyprotein or subgenomic replicons. Complete inhibition of RdRp activity in vitro was observed at equimolar concentrations of NS5B and mAb 5B-12B7, whereas RdRp activities of classica…
Viral and cellular determinants of hepatitis C virus RNA replication in cell culture.
2003
Studies on the replication of hepatitis C virus (HCV) have been facilitated by the development of selectable subgenomic replicons replicating in the human hepatoma cell line Huh-7 at a surprisingly high level. Analysis of the replicon population in selected cells revealed the occurrence of cell culture-adaptive mutations that enhance RNA replication substantially. To gain a better understanding of HCV cell culture adaptation, we characterized conserved mutations identified by sequence analysis of 26 independent replicon cell clones for their effect on RNA replication. Mutations enhancing replication were found in nearly every nonstructural (NS) protein, and they could be subdivided into at …
Biochemical properties of hepatitis C virus NS5B RNA-dependent RNA polymerase and identification of amino acid sequence motifs essential for enzymati…
1997
The NS5B protein of the hepatitis C virus (HCV) is an RNA-dependent RNA polymerase (RdRp) (S.-E. Behrens, L. Tomei, and R. De Francesco, EMBO J. 15:12-22, 1996) that is assumed to be required for replication of the viral genome. To further study the biochemical and structural properties of this enzyme, an NS5B-hexahistidine fusion protein was expressed with recombinant baculoviruses in insect cells and purified to near homogeneity. The enzyme was found to have a primer-dependent RdRp activity that was able to copy a complete in vitro-transcribed HCV genome in the absence of additional viral or cellular factors. Filter binding assays and competition experiments showed that the purified enzym…