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Nuclear Magnetic Resonance Analysis (nuclear + magnetic_resonance_analysis)
Selected AbstractsStructural and Biophysical Characterization of XIAP BIR3 G306E Mutant: Insights in Protein Dynamics and Application for Fragment-Based Drug DesignCHEMICAL BIOLOGY & DRUG DESIGN, Issue 3 2009Cathy D. Moore Previous reports describe modulators of X-linked inhibitor of apoptosis (XIAP),caspase interaction designed from the AVPI N-terminal peptide sequence of second mitochondria-derived activator of caspase. A fragment-based drug design strategy was initiated to identify therapeutic non-peptidomimetic antagonists of X-linked inhibitor of apoptosis protein,protein interactions. Fragments that bind to the AVPI binding site of BIR3 (bacculoviral inhibitory repeat) were identified, and to further localize the fragment binding within the AVPI binding site, a point mutation was designed which alters the dynamics of flexible loops and blocks PI region of the binding cleft, thus enabling definition of weakly bound small molecules in the AV portion of the binding cleft. Nuclear magnetic resonance analysis confirmed the G306E mutation stabilizes the AV pocket. Biophysical characterization of the mutant confirms conformation change within the PI sub-pocket as evidenced by a significant diminishment in binding affinity of AVPI mimetics, yet the binding affinity of the smaller AV mimetics is maintained or slightly improved in the mutant compared with wild-type. Additional data from non-covalent mass spectrometry analysis shows enhanced binding of AV mimetics to the G306E mutant over the wild-type. The presented data outline a protein engineering strategy that allowed mapping of AV-replacements with better sensitivity and precision. [source] Water-soluble, thermoresponsive, hyperbranched copolymers based on PEG-methacrylates: Synthesis, characterization, and LCST behaviorJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 13 2010Mario Luzon Abstract A series of water-soluble thermoresponsive hyperbranched copoly(oligoethylene glycol)s were synthesized by copolymerization of di(ethylene glycol) methacrylate (DEG-MA) and oligo(ethylene glycol) methacrylate (OEG-MA, Mw = 475 g/mol), with ethylene glycol dimethacrylate (EGD-MA) used as the crosslinker, via reversible addition fragmentation chain transfer polymerization. Polymers were characterized by size exclusion chromatography and nuclear magnetic resonance analyses. According to the monomer composition, that is, the ratio of OEG-MA: DEG-MA: EGD-MA, the lower critical solution temperature (LCST) could be tuned from 25 °C to 90 °C. The thermoresponsive properties of these hyperbranched copolymers were studied carefully and compared with their linear analogs. It was found that molecular architecture influences thermoresponsive behavior, with a decrease of around 5,10 °C in the LCST of the hyperbranched polymers compared with the LCST of linear chains. © 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 48: 2783,2792, 2010 [source] Synthesis and properties of biomimetic poly(L -glutamate)- b -poly(2-acryloyloxyethyllactoside)- b -poly(L -glutamate) triblock copolymersJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 22 2004Chang-Ming Dong Abstract A novel class of biomimetic glycopolymer,polypeptide triblock copolymers [poly(L -glutamate),poly(2-acryloyloxyethyllactoside),poly(L -glutamate)] was synthesized by the sequential atom transfer radical polymerization of a protected lactose-based glycomonomer and the ring-opening polymerization of ,-benzyl- L -glutamate N -carboxyanhydride. Gel permeation chromatography and nuclear magnetic resonance analyses demonstrated that triblock copolymers with defined architectures, controlled molecular weights, and low polydispersities were successfully obtained. Fourier transform infrared spectroscopy of the triblock copolymers revealed that the ,-helix/,-sheet ratio increased with the poly(benzyl- L -glutamate) block length. Furthermore, the water-soluble triblock copolymers self-assembled into lactose-installed polymeric aggregates; this was investigated with the hydrophobic dye solubilization method and ultraviolet,visible analysis. Notably, this kind of aggregate may be useful as an artificial polyvalent ligand in the investigation of carbohydrate,protein recognition and for the design of site-specific drug-delivery systems. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 5754,5765, 2004 [source] Characterization of a novel NCAM ligand with a stimulatory effect on neurite outgrowth identified by screening a combinatorial peptide libraryEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2002Lars C. B. Rønn Abstract The neural cell adhesion molecule, NCAM, plays a key role in neural development and plasticity mediating cell adhesion and signal transduction. By screening a combinatorial library of synthetic peptides with NCAM purified from postnatal day 10 rat brains, we identified a nonapeptide, termed NCAM binding peptide 10 (NBP10) and showed by nuclear magnetic resonance analysis that it bound the NCAM IgI module of NCAM. NBP10 modulated cell aggregation as well as neurite outgrowth induced specifically by homophilic NCAM binding. Moreover, both monomeric and multimeric forms of NBP10 stimulated neurite outgrowth from primary hippocampal neurons. The neurite outgrowth response to NBP10 was inhibited by a number of compounds previously shown to inhibit neurite outgrowth induced by homophilic NCAM binding, including voltage-dependent calcium channel antagonists, suggesting that NBP10 induced neurite outgrowth by activating a signal transduction pathway similar to that activated by NCAM itself. Moreover, an inhibitor of intracellular calcium mobilization, TMB-8, prevented NBP10-induced neurite outgrowth suggesting that NCAM-dependent neurite outgrowth also requires mobilization of calcium from intracellular calcium stores in addition to calcium influx from extracellular sources. By single-cell calcium imaging we further demonstrated that NBP10 was capable of inducing an increase in intracellular calcium in PC12E2 cells. Thus, the NBP10 peptide is a new tool for the study of molecular mechanisms underlying NCAM-dependent signal transduction and neurite outgrowth, and could prove to be a useful modulator of regenerative processes in the peripheral and central nervous system. [source] Preferential phosphorus leaching from an irrigated grassland soilEUROPEAN JOURNAL OF SOIL SCIENCE, Issue 2 2005G. S. Toor Summary Intact lysimeters (50 cm diameter, 70 cm deep) of silt loam soil under permanent grassland were used to investigate preferential transport of phosphorus (P) by leaching immediately after application of dairy effluent. Four treatments that received mineral P fertilizer alone (superphosphate at 45 kg P ha,1 year,1) or in combination with effluent (at , 40,80 kg P ha,1 year,1) over 2 years were monitored. Losses of total P from the combined P fertilizer and effluent treatments were 1.6,2.3 kg ha,1 (60% of overall loss) during eight drainage events following effluent application. The rest of the P lost (40% of overall loss) occurred during 43 drainage events following a significant rainfall or irrigation compared with 0.30 kg ha,1 from mineral P fertilizer alone. Reactive forms of P (mainly dissolved reactive P: 38,76%) were the dominant fractions in effluent compared with unreactive P forms (mainly particulate unreactive P: 15,56%). In contrast, in leachate following effluent application, particulate unreactive P was the major fraction (71,79%) compared with dissolved reactive P (1,7%). The results were corroborated by 31P nuclear magnetic resonance analysis, which showed that inorganic orthophosphate was the predominant P fraction present in the effluent (86%), while orthophosphate monoesters and diesters together comprised up to 88% of P in leachate. This shows that unreactive P forms were selectively transported through soil because of their greater mobility as monoesters (labile monoester P and inositol hexakisphosphate) and diesters. The short-term strategies for reducing loss of P after application of dairy effluent application should involve increasing the residence time of applied effluent in the soil profile. This can be achieved by applying effluent frequently in small amounts. [source] Effect of Guava (Psidium guajava L.) Leaf Extract on Glucose Uptake in Rat HepatocytesJOURNAL OF FOOD SCIENCE, Issue 5 2009Fang-Chi Cheng ABSTRACT:, People in oriental countries, including Japan and Taiwan, boil guava leaves (Psidium guajava L.) in water and drink the extract as a folk medicine for diabetes. The present study investigated the enhancement of aqueous guava leaf extract on glucose uptake in rat clone 9 hepatocytes and searched for the active compound. The extract was eluted with MeOH-H2O solutions through Diaion, Sephadex, and MCI-gel columns to separate into fractions with different polarities. The uptake test of 2-[1- 14C] deoxy-D-glucose in rat clone 9 hepatocytes was performed to evaluate the hypoglycemic effect of these fractions. The active compound was identified by nuclear magnetic resonance analysis and high-performance liquid chromatography (HPLC). The results revealed that phenolics are the principal component of the extract, that high polarity fractions of the guava leaf extract are enhancers to glucose uptake in rat clone 9 hepatocytes, and that quercetin is the major active compound. We suggest that quercetin in the aqueous extract of guava leaves promotes glucose uptake in liver cells, and contributes to the alleviation of hypoglycemia in diabetes as a consequence. [source] Sterol Composition of Pneumocystis jirovecii with Blocked 14,-Demethylase ActivityTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 6 2004JOSÉ-LUIS GINER ABSTRACT Several drugs that interact with membrane sterols or inhibit their syntheses are effective in clearing a number of fungal infections. The AIDS-associated lung infection caused by Pneumocystis jirovecii is not cleared by many of these therapies. Pneumocystis normally synthesizes distinct C28 and C29 24-alkylsterols, but ergosterol, the major fungal sterol, is not among them. Two distinct sterol compositional phenotypes were previously observed in P. jirovecii. One was characterized by ,7 C28 and C29 24-alkylsterols with only low proportions of higher molecular mass components. In contrast, the other type was dominated by high C31 and C32 24-alkylsterols, especially pneumocysterol. In the present study, 28 molecular species were elucidated by nuclear magnetic resonance analysis of a human lung specimen containing P. jirovecii representing the latter sterol profile phenotype. Fifteen of the 28 had the methyl group at C-14 of the sterol nucleus and these represented 96% of the total sterol mass in the specimen (excluding cholesterol). These results strongly suggest that sterol 14,-demethylase was blocked in these organisms. Twenty-four of the 28 were 24-alkylsterols, indicating that methylation of the C-24 position of the sterol side chain by S-adenosyl-L-methionine:sterol C-24 methyl transferase was fully functional. [source] | ||||||||||