Biomedical subjects

Roberto Consonni

Publications and source records attributed to Roberto Consonni.

5 recordsLinked to original sources

PubMed2004

1H NMR studies on Italian balsamic and traditional balsamic vinegars.

In the present work Principal Component Analysis applied to (1)H NMR spectra of balsamic and traditional balsamic vinegars is used to establish a simple and rapid aging determination protocol. Chemical composition of vinegar is dominated by carbohydrates even though several small components can be clearly observed in the proton NMR spectrum. Quantitative determination of some selected metabolites such as ethanol, acetic acid, malic acid, glucose, and HMF, considered as potential aging indicators, has been performed. (1)H NMR spectroscopy provides noninvasive characterization of such compounds, and our data demonstrate the validity of this approach, giving very promising results for assessing the quality of the final product.

Acetic Acid↗

PubMed2004

Structural characterization of imazalil/beta-cyclodextrin inclusion complex.

An equimolar inclusion complex between imazalil, a selected fungicide, and beta-cyclodextrin using an aqueous standard solution procedure has been obtained. The complex has been investigated in solution by (1)H and (13)C NMR techniques in combination with computational methods in order to establish a valuable analytical protocol through which to gain insight into the interactions of the inclusion complex in aqueous solution. Intramolecular NMR distance constraints have been detected and used for three-dimensional complex structure determination.

Cyclodextrins↗

PubMed2003

Structure determination and dynamics of peptides overlapping the catalytic hairpin of the Ras-specific GEF Cdc25(Mm).

Ras proteins are small G proteins playing a major role in eukaryotic signal transduction. Guanine nucleotide exchange factors (GEF) stimulate GDP/GTP exchange, resulting in the formation of the active Ras-GTP complex. In mammalian cells, two major Ras-specific GEF exist: Sos-like and Cdc25-like. To date, structural data are available only for Cdc25(Mm). We designed and synthesized Cdc25(Mm)-derived peptides spanning residues corresponding to the hSos1 HI helical hairpin that has been implicated in the GEF catalytic mechanism. NMR experiments on a chemically synthesized Cdc25(Mm)(1178-1222) peptide proved that helix I readily reaches a conformation very similar to the corresponding helix in hSos1, while residues corresponding to helix H in hSos1 show higher conformational flexibility. Molecular dynamics studies with the appropriate solvent model showed that different conformational spaces are available for the peptide. Since helix H is making several contacts with Ras and a Cdc25(Mm)(1178-1222) peptide is able to bind nucleotide-free Ras in a BIAcore assay, the peptide must be able to obtain the proper Ras-interacting conformation, at least transiently. These results indicate that rational design and improvement of the Ras-interacting peptides should take into account conformational and flexibility features to obtain molecules with the appropriate biochemical properties.

Amino Acid Sequence↗

PubMed2003

Investigations of Sso7d catalytic residues by NMR titration shifts and electrostatic calculations.

Sso7d is a small basic protein consisting of 62 amino acids isolated from the thermoacidophilic archeobacterium Sulfolobus solfataricus. The protein is endowed with DNA binding properties, RNase activity, and the capability of rescuing aggregated proteins in the presence of ATP. In this study, the electrostatic properties of Sso7d are investigated by using the Poisson-Boltzmann calculation of the surface potential distribution and following by NMR spectroscopy the proton chemical shift pH titration of acidic residues. Although the details of the catalytic mechanism still have to be defined, the results from NMR experiments confirm the possible involvement of Glu35 as the proton acceptor in the catalytic reaction, as seen by its abnormally high pK(a) value. Poisson-Boltzmann calculations and NMR titration shifts suggest the presence of a possible hydrogen bond between Glu35 and Tyr33, with a consequent rather rigid arrangement at these positions. Comparison with RNase T1 suggests that Tyr7 may be a good candidate for acting as a proton donor in the active site of Sso7d as shown by its low phenolic pK(a) of approximately 9.3. Titration experiments performed with the UpA, a RNA dinucleotide model, showed that the protein residues affected by the interaction are mainly located in a different region with respect to the surface affected by DNA recognition, in good agreement with the surface potential distribution found with electrostatic calculations.

Archaeal Proteins↗

PubMed2002

Separation of DNA fragments in hydroxylated poly(dimethylacrylamide) copolymers.

The novel polymer matrices reported here are low-viscosity sieving media for DNA capillary electrophoresis. This new family of matrices comprises copolymers of N,N-dimethylacrylamide with different monomers which increase polymer hydrophilicity. All these new copolymers self-coat on fused-silica capillaries. Resolution, peak spacing and peak width were the parameters taken into account to assess the influence of polymer structure on separation selectivity and efficiency. This work demonstrates that the performance of polydimethylacrylamide (PDMA) can be improved through copolymerization with hydrophilic monomers. The improvement is related to the efficiency parameter. The new copolymers, due to their low viscosity high sieving capacity and ability to suppress EOF, represent a better alternative to PDMA and are suitable replaceable matrices for capillary and microchip electrophoresis.

Acrylamides↗