Catedra de Bioquimica Basica de Macromoleculas

Santa Fe de la Vera Cruz, Argentina

Catedra de Bioquimica Basica de Macromoleculas

Santa Fe de la Vera Cruz, Argentina
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Deiber J.A.,CONICET | Piaggio M.V.,Catedra de Bioquimica Basica de Macromoleculas | Peirotti M.B.,CONICET
Electrophoresis | Year: 2013

This work explores the possibility of using the electrically charged "spherical soft particle" (SSP) to model the electrophoretic mobility of proteins in the low charge regime. The general framework concerning the electrophoretic mobility of the SSP already presented in the literature is analyzed and discussed here in particular for polyampholyte-polypeptide chains. In this regard, this theory is applied to BSA for different protocol pH values. The physicochemical conditions required to model proteins as SSP from their experimentally determined electrophoretic mobilities are established. In particular, the protein charge regulation phenomenon and the SSP particle core are included to study BSA having isoelectric point pI ≈ 5.71, within a wide range of bulk pH values. The results of this case study are compared with previous ones concerning the spherical porous particle and the spherical hard particle with occluded water. A discussion of chain conformations in the SSP polyampholyte layer is presented through estimations of the packing and friction fractal dimensions. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Deiber J.A.,CONICET | Piaggio M.V.,Catedra de Bioquimica Basica de Macromoleculas | Peirotti M.B.,CONICET
Electrophoresis | Year: 2013

This work explores the possibility of using the electrically charged "spherical porous particle" (SPP) to model the electrophoretic mobility of proteins in the low charge regime. In this regard, the electrophoretic mobility expression of the charged SPP (Hermans-Fujita model) is used and applied here to BSA and staphylococcal nuclease for different protocol pH values. The SPP is presented within the general framework of the "spherical soft particle" as described in the literature. The physicochemical conditions required to model proteins as SPP from their experimentally determined electrophoretic mobilities are established. It is shown that particle permeability and porosity and chain packing and friction fractal dimensions are relevant structural properties of proteins when hydrodynamic interaction between amino acid residues is present. The charge regulation phenomenon of BSA and staphylococcal nuclease with pIs ≈ 5.71 and 9.63, respectively, is described through the SPP within a wide range of bulk pH values. These case studies illustrate when the average regulating (pH) of the protein domain is lower and higher than the protocol pH. Further research for using the general spherical soft particle is also proposed on the basis of results and main conclusions. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Deiber J.A.,CONICET | Piaggio M.V.,Catedra de Bioquimica Basica de Macromoleculas | Peirotti M.B.,CONICET
Journal of Separation Science | Year: 2014

Neuronal activity loss may be due to toxicity caused by amyloid-beta peptides forming soluble oligomers. Here amyloid-beta peptides (1-42, 1-40, 1-39, 1-38, and 1-37) are characterized through the modeling of their experimental effective electrophoretic mobilities determined by a capillary zone electrophoresis method as reported in the literature. The resulting electrokinetic and hydrodynamic global properties are used to evaluate amyloid-beta peptide propensities to aggregation through pair particles interaction potentials and Brownian aggregation kinetic theories. Two background electrolytes are considered at 25°C, one for pH 9 and ionic strength I = 40 mM (aggregation is inhibited through NH4OH) the other for pH 10 and I = 100 mM (without NH4OH). Physical explanations of peptide oligomerization mechanisms are provided. The effect of hydration, electrostatic, and dispersion forces in the amyloidogenic process of amyloid-beta peptides (1-40 and 1-42) are quantitatively presented. The interplay among effective charge number, hydration, and conformation of chains is described. It is shown that amyloid-beta peptides (1-40 and 1-42) at pH 10, I = 100mM and 25°C, may form soluble oligomers, mainly of order 2 and 4, after an incubation of 48 h, which at higher times evolve and end up in complex structures (protofibrils and fibrils) found in plaques associated with Alzheimer's disease. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim


Deiber J.A.,CONICET | Piaggio M.V.,Catedra de Bioquimica Basica de Macromoleculas | Peirotti M.B.,CONICET
Electrophoresis | Year: 2011

Estimations of protein global conformations in well-specified physicochemical microenvironments are obtained through global structural parameters defined from polypeptide-scale analyses. For this purpose protein electrophoretic mobility data must be interpreted through a physicochemical CZE model to obtain estimates of protein equivalent hydrodynamic radius, effective and total charge numbers, hydration, actual ionizing pK and pH-near molecule. The electrical permittivity of protein domain is also required. In this framework, the solvent drag on proteins is obtained via the characteristic friction power coefficient associated with the number of amino acid residues defining the global chain conformation in solution. Also, the packing dimension related to the spatial distribution of amino acid residues within the protein domain is evaluated and discussed. These scaling coefficients together with the effective and total charge number fractions of proteins provide relevant interpretations of protein global conformations mainly from collapsed globule to hybrid chain regimes. Also, protein transport properties may be estimated within this framework. In this regard, the central role played by the friction power coefficient in the evaluation of these properties is highlighted. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Deiber J.A.,CONICET | Peirotti M.B.,CONICET | Piaggio M.V.,Catedra de Bioquimica Basica de Macromoleculas
Electrophoresis | Year: 2015

Neuronal activity loss may be due to toxicity caused mainly by amyloid-beta (1-40) and (1-42) peptides forming soluble oligomers. Here the amyloid-beta (12-28) peptide fragment (monomer) and its dimer are characterized at low pH through the modeling of their diffusion coefficients and effective electrophoretic mobilities. Translational diffusion coefficient experimental values of monomer and dimer analogs of this peptide fragment and monomer and dimer mixtures at thermodynamic equilibrium are used as reported in the literature for different monomer initial concentrations. The resulting electrokinetic and hydrodynamic global properties are employed to evaluate the amyloid-beta (12-28) peptide fragment propensity to dimerization through a thermodynamic theoretical framework. Therefore equilibrium constants are considered at pH 2.9 to elucidate one of the amyloidogenic mechanisms involving the central hydrophobic region LVFFA of the peptide spanning residues 17-21 associated with phenylalanine at positions 19 and 20 in the amino acid sequence of amyloid-beta peptides. An analysis demonstrating that peptide aggregation is a concentration-dependent process is provided, where both pair and intraparticle charge regulation phenomena become relevant. It is shown that the modeling of the effective electrophoretic mobility of the amyloid-beta (12-28) peptide fragment is crucial to understand the effect of hydrophobic region LVFFA in the amyloidogenic process. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Deiber J.A.,CONICET | Piaggio M.V.,Catedra de Bioquimica Basica de Macromoleculas | Peirotti M.B.,CONICET
Electrophoresis | Year: 2014

Several global chain properties of relatively long peptides composed of 20 amino acid residues are estimated through the modeling of their experimental effective electrophoretic mobilities determined by CZE for 2 < pH < 6. In this regard, an all l-α-eicosapeptide, including a secondary α-helix (Peptide 1) and its all retro d-inverso-α-eicosapeptide (Peptide 2), are considered. Despite Peptides 1 and 2 are isomeric chains, they do not present similar global conformations in the whole range of pH studied. These peptides may also differ in the quality of BGE components chain interactions depending on the pH value. Three Peptide 1 fragments (Peptides 3, 4, and 5) are also analyzed in this framework with the following purposes: (i) visualization of the effects of initial and final strands at each side of the α-helix on the global chain conformations of Peptide 1 at different pHs and (ii) analysis of global chain conformations of Peptides 1 and 2, and Peptide 1 fragments in relation to their pI values. Also, the peptide maximum and minimum hydrations predicted by the model, compatible with experimental effective electrophoretic mobilities at different pHs, are quantified and discussed, and needs for further research concerning chain hydration are proposed. It is shown that CZE is a useful analytical tool for peptidomimetic designs and purposes. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Deiber J.A.,CONICET | Peirotti M.B.,CONICET | Piaggio M.V.,Catedra de Bioquimica Basica de Macromoleculas
Electrophoresis | Year: 2016

Electrophoretic mobilities of amyloid-beta (1-40) and (1-42) peptides and their aggregates are modeled to study the amyloidogenic pathway associated with Alzheimeŕs Disease. The near molecule pH generated by the intraparticle charge regulation phenomenon during the oligomerization of amyloid-beta (1-40) and (1-42) peptides is evaluated and discussed as a relevant mechanism supporting the "amyloid cascade hypothesis" proposed in the literature. A theoretical framework associated with the oligomerization of amyloid-beta peptides including simple scaling laws and the consideration of electrokinetic and hydrodynamic global properties of oligomers is presented. The central finding is the explanation of the near molecule pH change toward the pI when the oligomerization number increases. These results allow one to rationalize consecutive physical stages that validate the amyloid cascade hypothesis. Concluding remarks involving mainly the effects of pair and intraparticle charge regulation phenomena on the amyloidogenic pathway with some suggestions for future research are provided. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


PubMed | CONICET and Catedra de Bioquimica Basica de Macromoleculas
Type: Journal Article | Journal: Electrophoresis | Year: 2016

Electrophoretic mobilities of amyloid-beta (1-40) and (1-42) peptides and their aggregates are modeled to study the amyloidogenic pathway associated with Alzheimers Disease. The near molecule pH generated by the intraparticle charge regulation phenomenon during the oligomerization of amyloid-beta (1-40) and (1-42) peptides is evaluated and discussed as a relevant mechanism supporting the amyloid cascade hypothesis proposed in the literature. A theoretical framework associated with the oligomerization of amyloid-beta peptides including simple scaling laws and the consideration of electrokinetic and hydrodynamic global properties of oligomers is presented. The central finding is the explanation of the near molecule pH change toward the pI when the oligomerization number increases. These results allow one to rationalize consecutive physical stages that validate the amyloid cascade hypothesis. Concluding remarks involving mainly the effects of pair and intraparticle charge regulation phenomena on the amyloidogenic pathway with some suggestions for future research are provided.

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