Search results for "Guanosine"
showing 10 items of 171 documents
Muscarinic acetylcholine receptor trafficking in streptolysin O-permeabilized MDCK cells.
1996
We investigated the validity of streptolysin O (SLO)-permeabilized Madin-Darbin canine kidney (MDCK) cells which express muscarinic acetylcholine receptors (mAChRs) coupled to pertussis toxin-sensitive guanine nucleotide-binding proteins (G proteins) for the study of the molecular machinery that regulated mAChR internalization and recycling. Exposure of SLO-permeabilized cells to carbachol-reduced cell surface receptor number by up to 40% without changing total receptor number. The kinetics and maximal extent of receptor internalization as well as the potency of carbachol to induce receptor internalization were almost identical in SLO-permeabilized and non-permeabilized cells. Using this se…
Adenylate storage, metabolism and utilization in coelomic cells of the polychaeteNereis virens (Annelida, polychaeta)
1996
Eleocytes are specialized coelomic cells in nereid annelids which assume a central role during germ cell development. They may contain extremely high concentrations of both adenosine monophosphate (AMP) and adenosine diphosphate (ADP) (each >10 μmol/ml of cell vol.), whereas the adenosine triphosphate (ATP) content is comparatively low (0.8 μmol/ml cell vol.).31P nuclear magnetic, resonance (NMR) studies of living eleocytes suggest the compartmentalization of both AMP and ADP in the large acidic vacuole characteristic for this cell type. Eleocytes are thus capable of storing high concentrations of ADP and AMP without inhibiting energy metabolism, by sequestering these compounds in a separat…
Restoration of Clostridium difficile toxin-B-inhibited phospholipase D by phosphatidylinositol 4,5-bisphosphate.
1996
Receptor signalling to phospholipase D (PLD) in human embryonic kidney (HEK) cells stably expressing the m3 muscarinic acetylcholine receptor apparently involves Rho proteins. Since phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] has been recognized as an essential cofactor for PLD activity and since activated Rho proteins have been reported to stimulate the synthesis of PtdIns(4,5)P2, we studied whether in HEK cells PLD activity is regulated by PtdIns(4,5)P2 and, in particular, whether PtdIns(4,5)P2 can restore PLD activity inhibited by Clostridium difficile toxin B, which inactivates Rho proteins. Addition of MgATP to permeabilized HEK cells increased basal PLD activity and potentia…
A modified guanosine phosphoramidite for click functionalization of RNA on the sugar edge
2012
A propargyl containing guanosine phosphoramidite was synthesized and incorporated into siRNA, enabling click-ligation with an azido fluorophore onto the nucleobase sugar edge. Duplex stability was not affected by labeling at this new site, which allowed deconvolution of the effects of label, structure and attachment site on RNAi activity.
Photochemical and Photobiological Studies of a Furonaphthopyranone as a Benzo-spaced Psoralen Analog in Cell-free and Cellular DNA
1997
Photobiological activities of the benzo-spaced psoralen analog furonaphthopyranone 3 have been investigated in cell-free and cellular DNA. The molecular geometry parameters of 3 suggest that it should not form interstrand crosslinks with DNA. With cell-free DNA no evidence for crosslinking but also not for monoadduct formation was obtained; rather, the unnatural furocoumarin 3 induces oxidative DNA modifications under near-UVA irradiation. The enzymatic assay of the photosensitized damage in cell-free PM2 DNA revealed the significant formation of lesions sensitive to formamidopyrimidine DNA glycosylase (Fpg protein). In the photooxidation of calf thymus DNA by the furonaphthopyranone 3, 0.2…
Presence of phosphorylatedO-ribosyl-adenosine In T-ψ-stem of yeast methlonine initiator tRNA
1989
We report in this paper on isolation and characterization of two unknown nucleosides G* and [A*] located in the T-psi-stem of yeast methionine initiator tRNA, using the combined means of HPLC protocols, real time UV-absorption spectrum, and post-run mass spectrometry by electron impact or fast atom bombardment. The G* nucleoside in position 65 was identified as unmodified guanosine. The structure of the unknown [A*] in position 64 was characterized as an isomeric form of O-ribosyl-adenosine by comparison of its chromatographic, UV-spectral and mass spectrometric properties with those of authentic O-alpha-ribofuranosyl-(1"----2')-adenosine isolated from biosynthetic poly(adenosine diphosphat…
Stereoselective Metabolic Activation of Dibenzo[a,l]Pyrene in the Human Mammary Carcinoma Cell Line MCF-7 Results in Formation of (-)-antiand (+)-syn…
1996
Abstract Dibenzo[a,l]pyrene (DB[a,l]P) is an important polycyclic aromatic hydrocarbon because of possible human exposure and its exceptionally high carcinogenicity in rodents. We examined the metabolism of DB[a,l]P and the formation of DB[a,l]P-DNA adducts in the human mammary carcinoma cell line (MCF-7). Analysis of the DNA adducts by 33P-postlabeling, immobilized boronate chromatography, HPLC and TLC demonstrated that DB[a,l]P is stereoselectively metabolized to specific optical isomers of DB[a,l]P-11,12-diol-13,14-epoxide (DB[a,l]PDE). The major anti-DB[a,l]PDE adduct formed in DB[a,l]P-treated MCF-7 cells resulted from reaction of (-)-anti-DB[a,l]PDE with DNA whereas the two major syn-…
Spectroscopic Studies of Oligonucleotide Adducts and Base Sequence Preference of Adducts Formed by the Stereoisomers of 7,8-Dihydroxy-9,10-epoxy-7,8,…
1996
Abstract 5′-d(CCTATAGATATCC) has been reacted with BPDE and the adducts derived from binding of BPDE to the exocyclic amino group of deoxyguanosine (dG) were studied with spectroscopic methods. The major dG-adducts of (+)- and (-)-anti-BPDE and a minor adduct of (+)-syn-BPDE showed the characteristics of trans-adducts. The major products formed with (+)- and (-)-syn-BPDE exhibit cis-adduct characteristics. Annealing of BPDE-modified oligonucleotides to complementary or partially complementary strands results in reduced fluorescence intensity in several cases and in others the intensity is markedly increased. These differences demonstrate that the adduct microenvironment is strongly influenc…
Changes in the proteome of sea urchin Paracentrotus lividus coelomocytes in response to LPS injection into the body cavity.
2020
Background The immune system of echinoderm sea urchins is characterised by a high degree of complexity that is not completely understood. The Mediterranean sea urchin Paracentrotus lividus coelomocytes mediate immune responses through phagocytosis, encapsulation of non-self particles, and production of diffusible factors including antimicrobial molecules. Details of these processes, and molecular pathways driving these mechanisms, are still to be fully elucidated. Principal findings In the present study we treated the sea urchin P. lividus with the bacterial lipopolysaccharide (LPS) and collected coelomocytes at different time-points (1, 3, 6 and 24 hours). We have shown, using label-free q…
Hydrogen bonding patterns of 7,9-dimethylguanine and its transplatinum(II) complexes
2002
Methylation at the N7 position is one of the most frequently naturally occurring modifications of guanosine. This alteration drastically changes the hydrogen bonding and acid–base properties of the guanine nucleobase. Here we show on the example of the model nucleobase 7,9-dimethylguanine that due to blockage of N7 of the purine ring, new hydrogen bonding patterns occur on the minor groove binding face of this nucleobase involving the ring nitrogen N3 and the exocyclic amino group N2H2. The free 7,9-dimethylguaninium ion and several transplatinum(II) complexes of the this ligand are presented and discussed. Methylation at N7 drastically changes the acid–base and hydrogen bonding properties …