ABSTRACT
Amyloid plaques are one of the two hallmarks of Alzheimer's disease (AD). They consist mainly of fibrils made of self-assembled amyloid-ß (Aß) peptides. Aß is produced in healthy brains from proteolytic cleavage of the amyloid precursor protein. Aß aggregates, in particular smaller, soluble aggregates, are toxic to cells. Hence, modulating the self-assembly of Aß became a very active field of research, with the aim to reduce the amount of the toxic aggregates of Aß or to block their toxic action. A great variety of molecules, chemical and biological, are able to modify the aggregation of Aß. Here we give an overview of the different mechanistic ways to modulate Aß aggregation and on which step in the self-assembly molecules can interfere. We discuss the aggregation modulators according to different important parameters, including the type of interaction (weak interaction, coordination or covalent bonds), the importance of kinetics and thermodynamics, the size of the modulating molecules, and binding specificity.
Subject(s)
Amyloid beta-Peptides/chemistry , Protein Aggregation, Pathological , Alzheimer Disease/metabolism , Alzheimer Disease/pathology , Amyloid beta-Peptides/metabolism , Animals , Humans , Plaque, Amyloid/metabolism , Plaque, Amyloid/pathology , Protein Aggregation, Pathological/metabolism , Protein Aggregation, Pathological/pathologyABSTRACT
SecB chaperones assist protein export in bacteria. However, certain SecB family members have diverged to become specialized toward the control of toxin-antitoxin (TA) systems known to promote bacterial adaptation to stress and persistence. In such tripartite TA-chaperone (TAC) systems, the chaperone was shown to assist folding and to prevent degradation of its cognate antitoxin, thus facilitating inhibition of the toxin. Here, we used both the export chaperone SecB of Escherichia coli and the tripartite TAC system of Mycobacterium tuberculosis as a model to investigate how generic chaperones can specialize toward the control of TA systems. Through directed evolution of SecB, we have identified and characterized mutations that specifically improve the ability of SecB to control our model TA system without affecting its function in protein export. Such a remarkable plasticity of SecB chaperone function suggests that its substrate binding surface can be readily remodeled to accommodate specific clients.
Subject(s)
Bacterial Proteins/chemistry , Escherichia coli/genetics , Gene Expression Regulation, Bacterial , Molecular Chaperones/chemistry , Mycobacterium tuberculosis/genetics , Toxin-Antitoxin Systems/genetics , Amino Acid Sequence , Amino Acid Substitution , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Binding Sites , Cloning, Molecular , Directed Molecular Evolution , Escherichia coli/metabolism , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Kinetics , Models, Molecular , Molecular Chaperones/genetics , Molecular Chaperones/metabolism , Mutagenesis, Site-Directed , Mutation , Mycobacterium tuberculosis/metabolism , Protein Binding , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Substrate SpecificityABSTRACT
Bacterial toxin-antitoxin (TA) systems, in which a labile antitoxin binds and inhibits the toxin, can promote adaptation and persistence by modulating bacterial growth in response to stress. Some atypical TA systems, known as tripartite toxin-antitoxin-chaperone (TAC) modules, include a molecular chaperone that facilitates folding and protects the antitoxin from degradation. Here we use a TAC module from Mycobacterium tuberculosis as a model to investigate the molecular mechanisms by which classical TAs can become 'chaperone-addicted'. The chaperone specifically binds the antitoxin at a short carboxy-terminal sequence (chaperone addiction sequence, ChAD) that is not present in chaperone-independent antitoxins. In the absence of chaperone, the ChAD sequence destabilizes the antitoxin, thus preventing toxin inhibition. Chaperone-ChAD pairs can be transferred to classical TA systems or to unrelated proteins and render them chaperone-dependent. This mechanism might be used to optimize the expression and folding of heterologous proteins in bacterial hosts for biotechnological or medical purposes.
Subject(s)
Bacterial Proteins/metabolism , Bacterial Toxins/metabolism , Molecular Chaperones/metabolism , Mycobacterium tuberculosis/physiology , Toxin-Antitoxin Systems/physiology , Protein Folding , Recombinant Proteins/metabolismABSTRACT
Oxidative stress is considered as an important factor and an early event in the etiology of Alzheimer's disease (AD). Cu bound to the peptide amyloid-ß (Aß) is found in AD brains, and Cu-Aß could contribute to this oxidative stress, as it is able to produce in vitro H2O2 and HOË in the presence of oxygen and biological reducing agents such as ascorbate. The mechanism of Cu-Aß-catalyzed H2O2 production is however not known, although it was proposed that H2O2 is directly formed from O2 via a 2-electron process. Here, we implement an electrochemical setup and use the specificity of superoxide dismutase-1 (SOD1) to show, for the first time, that H2O2 production by Cu-Aß in the presence of ascorbate occurs mainly via a free O2Ë(-) intermediate. This finding radically changes the view on the catalytic mechanism of H2O2 production by Cu-Aß, and opens the possibility that Cu-Aß-catalyzed O2Ë(-) contributes to oxidative stress in AD, and hence may be of interest.