Bovine Serum Albumin protofibril-like aggregates formation: Solo but not simple mechanism

Vetri, Valeria; D’Amico, Michele; Foderà, Vito; Leone, Maurizio; Ponzoni, Andrea; Sberveglieri, G.; Militello, Valeria

doi:10.1016/j.abb.2011.01.024

Cited by 90 publications

(84 citation statements)

References 80 publications

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“…5-7.4 in various period of times [26,34,48]; it has also been observed that pH values close to the isoelectric point would result in more amorphous aggregates [34,48].…”

Section: Discussionmentioning

confidence: 91%

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Response Surface Methodology for Optimizing the Bovine Serum Albumin Fibrillation

et al. 2012

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Amyloid fibrils are considered as nanostructures that could be formed by ordered self-assembly of the partially-folded states of many different peptides or proteins. In this study, bovine serum albumin was used as a model protein whose ordered aggregation (fibrillation) was optimized. Response surface methodology (RSM) was used in a design that contained a total of 30 experimental trials. The first 24 were organized in a factorial design and from 25 to 30 involved the replications of the central points. Data obtained from RSM were subjected to the analysis of variance (ANOVA) and analyzed using a second order polynomial equation. Subsequent testing of the suggested experimental parameters was done in vitro with Congo red spectrophotometric assay. Protein concentration, pH, temperature and time of incubation were the variables used in this study. Responses were assessed by measuring absorbance in 540 nm (characteristic of amyloid formation) and maximal wavelength. Concomitant effects of variables were assessed in surface plots that each considered two of the variables. Interestingly, the pattern obtained by monitoring absorbance at 540 nm and absorbance in maximal wavelength were identical in most cases. We are reporting the optimum concentration of protein, pH, temperature and time at 5 mg ml⁻¹, 3.02 and 72 °C and 48 h, respectively. Our findings suggest that use of Congo red spectrophotometric test, as a simple and affordable assay could be suggested as a first test for assessing fibril formation of proteins. Absorbance in maximal wavelength is recommended as a significant indication of fibril formation.

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“…5-7.4 in various period of times [26,34,48]; it has also been observed that pH values close to the isoelectric point would result in more amorphous aggregates [34,48].…”

Section: Discussionmentioning

confidence: 91%

“…Bovine serum albumin (BSA) has been extensively used as a model to study protein aggregation [6,8,24,27,34,35,48,49,51] and fibrillation [10,48]. BSA is essentially an all a protein, which is transformed to b-form upon destabilizing conditions [23,24,36].…”

Section: Introductionmentioning

confidence: 99%

Response Surface Methodology for Optimizing the Bovine Serum Albumin Fibrillation

et al. 2012

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“…On the other hand, formation of amyloid structures, requiring extensive conformational changes in proteins (Vetri et al, 2007a) would be favored by conditions where inter-molecular hydrogen bonds occur preferentially over hydrogen bonds that form with the solvent (e.g. water), indicating that electrostatic and hydrophobic forces could be decisive factors in defining aggregate morphology (Vetri et al, 2011).…”

Section: Amorphous Aggregationmentioning

confidence: 99%

Heme binding site in apomyoglobin may be effectively targeted with small molecules to control aggregation

Azami-Movahed

Shariatizi

Sabbaghian

et al. 2013

The International Journal of Biochemistry & Cell Biology

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“…This is dictated by the high protein concentration necessary to permit the effective aggregation of partially misfolded proteins in order to form soluble protein oligomers. Previously, protein-only amyloid has been generated by treating native proteins with high temperature, hydrophobic solvent, denaturing agents, or extreme pH exposure [19][20][21][22] . However, soluble protein oligomers, if detectable at all, are only formed temporarily in solution with very limited abundance.…”

Section: Discussionmentioning

confidence: 99%

“…Previously, various methods have been developed to generate amyloid in vitro by applying conditions known to favor protein misfolding, such as high temperature, hydrophobic environment, and pH variations [19][20][21][22][23] . However, they associate with various problems, such as low efficiency, reversible folding, large batch variations or slow kinetics.…”

Section: Introductionmentioning

confidence: 99%

Rapid Generation of Amyloid from Native Proteins <em>In vitro</em>

Dorta‐Estremera

Cao

2013

JoVE

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Proteins carry out crucial tasks in organisms by exerting functions elicited from their specific three dimensional folds. Although the native structures of polypeptides fulfill many purposes, it is now recognized that most proteins can adopt an alternative assembly of beta-sheet rich amyloid. Insoluble amyloid fibrils are initially associated with multiple human ailments, but they are increasingly shown as functional players participating in various important cellular processes. In addition, amyloid deposited in patient tissues contains nonproteinaceous components, such as nucleic acids and glycosaminoglycans (GAGs). These cofactors can facilitate the formation of amyloid, resulting in the generation of different types of insoluble precipitates. By taking advantage of our understanding how proteins misfold via an intermediate stage of soluble amyloid precursor, we have devised a method to convert native proteins to amyloid fibrils in vitro. This approach allows one to prepare amyloid in large quantities, examine the properties of amyloid generated from specific proteins, and evaluate the structural changes accompanying the conversion. Video LinkThe video component of this article can be found at

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Bovine Serum Albumin protofibril-like aggregates formation: Solo but not simple mechanism

Cited by 90 publications

References 80 publications

Response Surface Methodology for Optimizing the Bovine Serum Albumin Fibrillation

Response Surface Methodology for Optimizing the Bovine Serum Albumin Fibrillation

Heme binding site in apomyoglobin may be effectively targeted with small molecules to control aggregation

Rapid Generation of Amyloid from Native Proteins <em>In vitro</em>

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