Concentration dependent switch in the kinetic pathway of lysozyme fibrillation: Spectroscopic and microscopic analysis

Kumar, E. K. Pramod; Prasad, Deepak Kumar; Prabhu, N. Prakash

doi:10.1016/j.saa.2017.04.011

Cited by 17 publications

(6 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Three independent incubation experiments with different ATs were performed for all measurements, which further ensured the reliability of the results. In reality, both the morphological changes of HEWL and the typical S-shaped curve with the lower AT were consistent with the results previously reported at room temperature. ,, …”

Section: Methodssupporting

confidence: 92%

“…In reality, both the morphological changes of HEWL and the typical S-shaped curve with the lower AT were consistent with the results previously reported at room temperature. 5,15,35 Measurement of Fiber Morphology by AFM. All the protein solution used in the AFM experiment was the 50-fold diluted supernatant with Milli-Q water after centrifugation.…”

Section: ■ Conclusionmentioning

confidence: 99%

“…The mature fibrils of HEWL possess rigidity, chemical stability, and high aspect ratio, which are well known for the design of enzymatically active hydrogel particles or drug carrier hydrogels. − Accordingly, the relevant results obtained from the study on HEWL are of reference value for most protein self-assembly research. , Considering the complexity and variability of the in vivo environment, abundant in vitro experiments connected with lysozyme fibrillation are being carried out. To date, many factors, such as concentration, solution temperature, pH, pressure, radiation, stirring rate, small molecules, and the surface or interface, have been identified to affect protein fibrillation in vitro. ,, Although these parameters have been controlled, it remains challenging to upward the precise repeatability of amyloid fibrillation kinetics between laboratories as amyloid fibrillation is a highly sensitive process, such as the inconsistent kinetic fluorescent curves and the quite stochastic mature fibrils. ,, Accordingly, the exact molecular mechanism of amyloid fibrillation is still under debate. This will be very detrimental to drug screening assays targeting the treatment of neurodegenerative diseases.…”

Section: Introductionmentioning

confidence: 99%

“…To date, many factors, such as concentration, solution temperature, pH, pressure, radiation, stirring rate, small molecules, and the surface or interface, have been identified to affect protein fibrillation in vitro. 4 , 14 , 15 Although these parameters have been controlled, it remains challenging to upward the precise repeatability of amyloid fibrillation kinetics between laboratories as amyloid fibrillation is a highly sensitive process, such as the inconsistent kinetic fluorescent curves and the quite stochastic mature fibrils. 4 , 16 , 17 Accordingly, the exact molecular mechanism of amyloid fibrillation is still under debate.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Ambient Temperature Affects Protein Self-Assembly by Interfering with the Interfacial Aggregation Behavior

et al. 2023

View full text Add to dashboard Cite

Amyloid fibrillation is known to be associated with degenerative diseases, and mature fibrils are also considered as valuable biomedical materials. Thus, the mechanism and influencing factors of fibrillation have always been the focus of research. However, in vitro studies are always plagued by low reproducibility of kinetics and the molecular mechanism of amyloid fibrillation is under debate until now. Here, we identified the ambient temperature (AT) as a non-negligible interfering factor in in vitro self-assembly of globular protein hen egg-white lysozyme for the first time. By multimodal molecular spectroscopy methods, not only the effect of ATs on the kinetics of protein aggregation was described but also the conformational changes of the molecular structure with different ATs were captured. Through investigating the dependence of interfacial area and catalysis, the reason for this influence was construed by the various aggregation behaviors of protein molecules in the two-phase interface. The results suggest that in vitro mechanism research on protein fibrillation needs to first clarify the AT for a more accurate comparative analysis. The proposal of this concept will provide a new clue for a deeper understanding of the mechanism of protein self-assembly and may have an impact on evaluating the efficiency of amyloid accelerators or inhibitors based on the comparative analysis of protein self-assembly.

show abstract

Section: Methodssupporting

confidence: 92%

Section: ■ Conclusionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Ambient Temperature Affects Protein Self-Assembly by Interfering with the Interfacial Aggregation Behavior

et al. 2023

View full text Add to dashboard Cite

show abstract

“…Model for nucleation-independent mechanism Some amyloid-forming proteins show a simple exponential growth without the observation of a nucleation phase [240][241][242][243]. Such a nucleation-independent pathway can also be accomplished by the addition of seeds or by changes in the protein concentration or the environment condition, as well as by the introduction of mutations in the protein [194,216,240].…”

Section: Models For Nucleation-dependent Mechanismsmentioning

confidence: 99%

A guide to studying protein aggregation

et al. 2021

View full text Add to dashboard Cite

Disrupted protein folding or decreased protein stability can lead to the accumulation of (partially) un-or misfolded proteins, which ultimately cause the formation of protein aggregates. Much of the interest in protein aggregation is associated with its involvement in a wide range of human diseases and the challenges it poses for large-scale biopharmaceutical manufacturing and formulation of therapeutic proteins and peptides. On the other hand, protein aggregates can also be functional, as observed in nature, which triggered its use in the development of biomaterials or therapeutics as well as for the improvement of food characteristics. Thus, unmasking the various steps involved in protein aggregation is critical to obtain a better understanding of the underlying mechanism of amyloid formation. This knowledge will allow a more tailored development of diagnostic methods and treatments for amyloid-associated diseases, as well as applications in the fields of new (bio)materials, food technology and therapeutics. However, the complex and dynamic nature of the aggregation process makes the study of protein aggregation challenging. To provide guidance on how to analyze protein aggregation, in this review we summarize the most commonly investigated aspects of protein aggregation with some popular corresponding methods.

show abstract

Amino Acid Functionalized Superparamagnetic Nanoparticles Inhibit Lysozyme Amyloid Fibrillization

Antošová

Bednarikova

Koneracká

et al. 2019

Chemistry A European J

View full text Add to dashboard Cite

Nanoparticles have great potential to be used in various biomedical applications, including therapy or diagnosis of amyloid‐related diseases. The physical and chemical properties of iron oxide superparamagnetic nanoparticles (MNPs) functionalized with different amino acids (AAs), namely, with lysine (Lys), glycine (Gly), or tryptophan (Trp), have been characterized. The cytotoxicity of nanoparticles and their effect on amyloid fibrillization of lysozymes in vitro was also verified. The AA‐MNPs under study are nontoxic to human SHSY5Y neuroblastoma cells. Moreover, the AA‐MNPs were able to significantly inhibit lysozyme amyloid fibrillization and destroy amyloid fibrils. Kinetic studies revealed that the presence of AA‐MNPs affected lysozyme fibrillization, namely, the lag phase and steady‐state phase of the growth curves. The most effective activities were observed for Trp‐MNPs, which revealed the importance of aromatic rings in the structure of AAs used as coating agents. The obtained results indicate the possible application of these AA‐MNPs in the treatment of amyloid diseases associated with lysozyme or other amyloidogenic proteins.

show abstract

Concentration dependent switch in the kinetic pathway of lysozyme fibrillation: Spectroscopic and microscopic analysis

Cited by 17 publications

References 54 publications

Ambient Temperature Affects Protein Self-Assembly by Interfering with the Interfacial Aggregation Behavior

Ambient Temperature Affects Protein Self-Assembly by Interfering with the Interfacial Aggregation Behavior

A guide to studying protein aggregation

Amino Acid Functionalized Superparamagnetic Nanoparticles Inhibit Lysozyme Amyloid Fibrillization

Contact Info

Product

Resources

About