Human Serum Albumin Aggregation/Fibrillation and its Abilities to Drugs Binding

Maciążek-Jurczyk, Małgorzata; Janas, Kamil; Pożycka, Jadwiga; Szkudlarek, Agnieszka; Rogóż, Wojciech; Owczarzy, Aleksandra; Kulig, Karolina

doi:10.3390/molecules25030618

Cited by 62 publications

(40 citation statements)

References 58 publications

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“…This approach was used in the evaluation of the self-assembly of the 33-mer peptide at different concentrations at it is observed in Figure 3 D, E. Here, a red-shift is observed indicating that the tyrosines of the peptide are located in a more hydrophobic environment upon aggregation [ 30 ]. Additionally, this approach was used to evaluate the aggregation upon cation binding of Azurin and RNAse T1 [ 58 ], human serum albumin aggregation into fibrils [ 59 ], gliadin self-organization in aqueous solutions [ 60 ] and the assembly of the Ac-Phe-Phe-Cys-NH 2 (Ac-FFC-NH 2 ) amyloid peptide model [ 61 ].…”

Section: Uv-vis Absorption Spectroscopy and Turbidity Are Initial mentioning

confidence: 99%

Evaluation of Peptide/Protein Self-Assembly and Aggregation by Spectroscopic Methods

2020

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The self-assembly of proteins is an essential process for a variety of cellular functions including cell respiration, mobility and division. On the other hand, protein or peptide misfolding and aggregation is related to the development of Parkinson’s disease and Alzheimer’s disease, among other aggregopathies. As a consequence, significant research efforts are directed towards the understanding of this process. In this review, we are focused on the use of UV-Visible Absorption Spectroscopy, Fluorescence Spectroscopy and Circular Dichroism to evaluate the self-organization of proteins and peptides in solution. These spectroscopic techniques are commonly available in most chemistry and biochemistry research laboratories, and together they are a powerful approach for initial as well as routine evaluation of protein and peptide self-assembly and aggregation under different environmental stimulus. Furthermore, these spectroscopic techniques are even suitable for studying complex systems like those in the food industry or pharmaceutical formulations, providing an overall idea of the folding, self-assembly, and aggregation processes, which is challenging to obtain with high-resolution methods. Here, we compiled and discussed selected examples, together with our results and those that helped us better to understand the process of protein and peptide aggregation. We put particular emphasis on the basic description of the methods as well as on the experimental considerations needed to obtain meaningful information, to help those who are just getting into this exciting area of research. Moreover, this review is particularly useful to those out of the field who would like to improve reproducibility in their cellular and biomedical experiments, especially while working with peptide and protein systems as an external stimulus. Our final aim is to show the power of these low-resolution techniques to improve our understanding of the self-assembly of peptides and proteins and translate this fundamental knowledge in biomedical research or food applications.

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Section: Uv-vis Absorption Spectroscopy and Turbidity Are Initial mentioning

confidence: 99%

Evaluation of Peptide/Protein Self-Assembly and Aggregation by Spectroscopic Methods

2020

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“…It has been shown that all α-helical proteins show a strong minimum at 222 nm and 208–210 nm and a strong maximum at 191–193 nm. In the case of β-sheet protein, a single negative band may be observed in the 210–225 nm wavelength range and a strong positive band in the 190–200 nm wavelength range, although their intensities are noticeably lower than those for α-proteins [ 66 ]. The CD spectra recorded for buffered solutions of insulin ( 4 ) and hot-spot fragments ( 5 ), ( 6 ) in the presence of HSA as well as for native insulin are presented in Figure 9 a,b.…”

Section: Resultsmentioning

confidence: 99%

Human Serum Albumin Binds Native Insulin and Aggregable Insulin Fragments and Inhibits Their Aggregation

et al. 2020

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The purpose of this study was to investigate whether Human Serum Albumin (HSA) can bind native human insulin and its A13–A19 and B12–B17 fragments, which are responsible for the aggregation of the whole hormone. To label the hormone and both hot spots, so that their binding positions within the HSA could be identified, 4-(1-pyrenyl)butyric acid was used as a fluorophore. Triazine coupling reagent was used to attach the 4-(1-pyrenyl)butyric acid to the N-terminus of the peptides. When attached to the peptides, the fluorophore showed extended fluorescence lifetimes in the excited state in the presence of HSA, compared to the samples in buffer solution. We also analyzed the interactions of unlabeled native insulin and its hot spots with HSA, using circular dichroism (CD), the microscale thermophoresis technique (MST), and three independent methods recommended for aggregating peptides. The CD spectra indicated increased amounts of the α-helical secondary structure in all analyzed samples after incubation. Moreover, for each of the two unlabeled hot spots, it was possible to determine the dissociation constant in the presence of HSA, as 14.4 µM (A13–A19) and 246 nM (B12–B17). Congo Red, Thioflavin T, and microscopy assays revealed significant differences between typical amyloids formed by the native hormone or its hot-spots and the secondary structures formed by the complexes of HSA with insulin and A13–A19 and B12–B17 fragments. All results show that the tested peptide-probe conjugates and their unlabeled analogues interact with HSA, which inhibits their aggregation.

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“…This allowed for a significant simplification of the procedure and reduced analysis time. The amount of unbound ketoprofen was determined in the supernatant using UV-Vis spectroscopy and the amount of protein-bound ketoprofen was calculated using Equation (1). Because the results obtained for the amount of unbound ketoprofen with free HSA by measuring absorbance and by HPLC were in agreement, for immobilized HSA, the results were based only on measurements of the absorption spectrum.…”

Section: Interaction Of Immobilized Hsa With Kpmentioning

confidence: 99%

“…This makes it one of the most frequently used NSAIDs. It is known that ketoprofen is bounded by the HSA structure at the Sudlow II site (Figure 2b), which, according to literature reports, is very sensitive to oxidizing agents [1]. The properties of KP and its wide application why in the study of the interaction of the drug-HSA it can be used as a model active substance.…”

Section: Introductionmentioning

confidence: 98%

“…Human serum albumin (HSA) is one of the most important globular plasma proteins containing a single heart-shaped polypeptide chain of 585 amino acid residues organized in three homologous domains (I, II and III) with two subdomains, A and B (Figure 1). The most important role of HSA is the ability to bind and transport substances [1]. Compounds bound and transported by albumin are fatty acids, bilirubin, metal ions, and, above all, drugs.…”

Section: Introductionmentioning

confidence: 99%

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Testing for Ketoprofen Binding to HSA Coated Magnetic Nanoparticles under Normal Conditions and after Oxidative Stress

et al. 2020

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Binding and transport of ligands is one of the most important functions of human blood serum proteins. Human serum albumin is found in plasma at the highest concentration. Because of this, it is important to study protein–drug interactions for this albumin. Since there is no single model describing this interaction, it is necessary to measure it for each active substance. Drug binding should also be studied in conditions that simulate pathological conditions of the body, i.e., after oxidative stress. Due to this, it is expected that the methods for testing these interactions need to be easy and fast. In this study, albumin immobilized on magnetic nanoparticles was successfully applied in the study of protein–drug binding. Ketoprofen was selected as a model drug and interactions were tested under normal conditions and artificially induced oxidative stress. The quality of obtained results for immobilized protein was confirmed with those for free albumin and literature data. It was shown that the type of magnetic core coverage does not affect the quality of the obtained results. In summary, a new, fast, effective, and universal method for testing protein–drug interactions was proposed, which can be performed in most laboratories.

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Human Serum Albumin Aggregation/Fibrillation and its Abilities to Drugs Binding

Cited by 62 publications

References 58 publications

Evaluation of Peptide/Protein Self-Assembly and Aggregation by Spectroscopic Methods

Evaluation of Peptide/Protein Self-Assembly and Aggregation by Spectroscopic Methods

Human Serum Albumin Binds Native Insulin and Aggregable Insulin Fragments and Inhibits Their Aggregation

Testing for Ketoprofen Binding to HSA Coated Magnetic Nanoparticles under Normal Conditions and after Oxidative Stress

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