Advances in recent nanoscience technologies have generated a new compilation of biocompatible, fluorescent nanoparticles derived from semiconductor quantum dots (QDs). QDs are extremely small in size and possess very large surface areas, which gives them unique physical properties and applications distinct from those of bulk systems. When exposed to biological fluid, these QDs may become coated with proteins and other biomolecules given their dynamic nature. These protein-QD systems may affect or enhance the changes in protein structure and stability, leading to the destruction of biological function. It is believed that these QDs can act as nucleation centers and subsequently promote protein fibril formation. Protein fibrillation is closely associated with many fatal human diseases, including neurodegenerative diseases and a variety of systemic amyloidoses. This topic of protein-QD interaction brings about many key issues and concerns, especially with respect to the potential risks to human health and the environment. Herein, the behavioral effects of dihydrolipoic acid (DHLA)-capped CdSe/ZnS (core/shell) QDs in hen egg-white lysozyme (HEWL) and human serum albumin (HSA) protein systems were systematically analyzed. This study gives rise to a better understanding of the potentially useful application of these protein-QD systems in nanobiotechnology and nanomedicine as a bioimaging tool and/or as a reference for controlled biological self-assembly processes.

This work investigates the influence of mutations at selected positions on the structure formation of the Alzheimer’s disease peptide amyloid β. Amyloid β is a member of the class of intrinsically disordered proteins that can aggregate into fibrils, which are characterized by a highly stable secondary structure, called cross-β structure. A central contact during fibrillation is the hydrophobic F19-L34 contact, which is located within the core of the cross-β structure. Modifications of this contact are known to influence the local molecular structure whereas the fibril morphology and the cross-β structure remain stable. In contrast, toxicity of amyloid β was completely lost for all previously investigated mutants of F19 and L34. This work characterizes the properties of this contact and answers the question what the minimally tolerated modifications are. To characterize the structure, structure formation process and biological activity of the Aβ variants a set of experiments was carried out. The local structure and dynamics were investigated using NMR experiments focusing on 13C-chemical shift changes and 1H-13C dipolar couplings, respectively. The fibril morphology and cross- β structure was verified by electron microscopy, circular dichroism spectroscopy and X-ray diffraction. Toxicity and biological activity was investigated using complementary cell culture experiments. The work was divided in three parts. First, L34 was substituted with three highly similar amino acids: the isomer isoleucine, valine that is one methylene group shorter but also a branched chain amino acid and the stereoisomer D-leucine. The L34 position proved to be important for the initiation of the structure formation, oligomer stability, fibril growth and the biological activity of amyloid β. These characteristics and properties were highly sensitive also to minor modifications but the different mutants showed no specific but qualitatively similar effects. The second part complemented previous mutation studies of the F19 position. Four new mutants were designed testing mild modification of the F19-L34 contact: phenylglycine and the homophenylalanine (S)-2-amino-4-phenyl-butyric acid change the length of the side chain, cyclohexyl-alanine eliminates the π-aromaticity of the ring system and increases the 3D steric demand, and (1-naphtyl)-alanine increases the 2D steric demand while maintaining the aromaticity. Mutations at the F19 position caused qualitatively similar effects as L34 modifications but proved to have quantitatively greater impact. Furthermore, they showed some specificity as steric constraints caused larger changes than modifications of the ring system. The third part investigates the influence of β-methylamino-L-alanine (BMAA) substitutions at positions F19, S8, and S26. The serine to BMAA substitutions were included because of their potential medical relevance. A F19BMAA substitution caused similar effects like other modifications at this position. Replacement of serine lead to a structural reorientation of the Aβ N-terminus and turn region. Furthermore, the pathways of the cell response changed from mitochondrial activity and plasma membrane integrity to apoptosis and neuronal stress reaction. Summarizing, it could be shown that, although the formation and structure of amyloid β fibrils is robust against different modifications the fibrillation kinetics, local structure and especially biological activity is highly sensitive and to some extend specific to even minor modifications.

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