459 research outputs found
Molecular crowding and RNA synergize to promote phase separation, microtubule interaction, and seeding of Tau condensates
Biomolecular condensation of the neuronal microtubule-associated protein Tau (MAPT) can be induced by coacervation with polyanions like RNA, or by molecular crowding. Tau condensates have been linked to both functional microtubule binding and pathological aggregation in neurodegenerative diseases. We find that molecular crowding and coacervation with RNA, two conditions likely coexisting in the cytosol, synergize to enable Tau condensation at physiological buffer conditions and to produce condensates with a strong affinity to charged surfaces. During condensate-mediated microtubule polymerization, their synergy enhances bundling and spatial arrangement of microtubules. We further show that different Tau condensates efficiently induce pathological Tau aggregates in cells, including accumulations at the nuclear envelope that correlate with nucleocytoplasmic transport deficits. Fluorescent lifetime imaging reveals different molecular packing densities of Tau in cellular accumulations and a condensate-like density for nuclear-envelope Tau. These findings suggest that a complex interplay between interaction partners, post-translational modifications, and molecular crowding regulates the formation and function of Tau condensates. Conditions leading to prolonged existence of Tau condensates may induce the formation of seeding-competent Tau and lead to distinct cellular Tau accumulations
Potent Tau Aggregation Inhibitor D-Peptides Selected against Tau-Repeat 2 Using Mirror Image Phage Display
Alzheimer's disease and other Tauopathies are associated with neurofibrillary tangles composed of Tau protein, as well as toxic Tau oligomers. Therefore, inhibitors of pathological Tau aggregation are potentially useful candidates for future therapies targeting Tauopathies. Two hexapeptides within Tau, designated PHF6* (275-VQIINK-280) and PHF6 (306-VQIVYK-311), are known to promote Tau aggregation. Recently, the PHF6* segment has been described as the more potent driver of Tau aggregation. We therefore employed mirror-image phage display with a large peptide library to identify PHF6* fibril binding peptides consisting of D-enantiomeric amino acids. The suitability of D-enantiomeric peptides for inā
vivo applications, which are protease stable and less immunogenic than L-peptides, has already been demonstrated. The identified D-enantiomeric peptide MMD3 and its retro-inverso form, designated MMD3rev, inhibited inā
vitro fibrillization of the PHF6* peptide, the repeat domain of Tau as well as full-length Tau. Dynamic light scattering, pelleting assays and atomic force microscopy demonstrated that MMD3 prevents the formation of tau Ī²-sheet-rich fibrils by diverting Tau into large amorphous aggregates. NMR data suggest that the D-enantiomeric peptides bound to Tau monomers with rather low affinity, but ELISA (enzyme-linked immunosorbent assay) data demonstrated binding to PHF6* and full length Tau fibrils. In addition, molecular insight into the binding mode of MMD3 to PHF6* fibrils were gained by in silico modelling. The identified PHF6*-targeting peptides were able to penetrate cells. The study establishes PHF6* fibril binding peptides consisting of D-enantiomeric amino acids as potential molecules for therapeutic and diagnostic applications in AD research
A novel D-amino acid peptide with therapeutic potential (ISAD1) inhibits aggregation of neurotoxic disease-relevant mutant Tau and prevents Tau toxicity in vitro
Background: Alzheimer's disease (AD), the most common form of dementia, is a progressive neurodegenerative disorder that mainly affects older adults. One of the pathological hallmarks of AD is abnormally aggregated Tau protein that forms fibrillar deposits in the brain. In AD, Tau pathology correlates strongly with clinical symptoms, cognitive dysfunction, and neuronal death.
Methods: We aimed to develop novel therapeutic D-amino acid peptides as Tau fibrillization inhibitors. It has been previously demonstrated that D-amino acid peptides are protease stable and less immunogenic than L-peptides, and these characteristics may render them suitable for in vivo applications. Using a phage display procedure against wild type full-length Tau (TauFL), we selected a novel Tau binding L-peptide and synthesized its D-amino acid version ISAD1 and its retro inversed form, ISAD1rev, respectively.
Results: While ISAD1rev inhibited Tau aggregation only moderately, ISAD1 bound to Tau in the aggregation-prone PHF6 region and inhibited fibrillization of TauFL, disease-associated mutant full-length Tau (TauFLĪK, TauFL-A152T, TauFL-P301L), and pro-aggregant repeat domain Tau mutant (TauRDĪK). ISAD1 and ISAD1rev induced the formation of large high molecular weight TauFL and TauRDĪK oligomers that lack proper Thioflavin-positive Ī²-sheet conformation even at lower concentrations. In silico modeling of ISAD1 Tau interaction at the PHF6 site revealed a binding mode similar to those known for other PHF6 binding peptides. Cell culture experiments demonstrated that ISAD1 and its inverse form are taken up by N2a-TauRDĪK cells efficiently and prevent cytotoxicity of externally added Tau fibrils as well as of internally expressed TauRDĪK.
Conclusions: ISAD1 and related peptides may be suitable for therapy development of AD by promoting off-pathway assembly of Tau, thus preventing its toxicity.
Keywords: Alzheimerās disease; D-amino acidĀ peptides; Phage display; Tau aggregation inhibitors; Therap
Transport by molecular motors in the presence of static defects
The transport by molecular motors along cytoskeletal filaments is studied
theoretically in the presence of static defects. The movements of single motors
are described as biased random walks along the filament as well as binding to
and unbinding from the filament. Three basic types of defects are
distinguished, which differ from normal filament sites only in one of the
motors' transition probabilities. Both stepping defects with a reduced
probability for forward steps and unbinding defects with an increased
probability for motor unbinding strongly reduce the velocities and the run
lengths of the motors with increasing defect density. For transport by single
motors, binding defects with a reduced probability for motor binding have a
relatively small effect on the transport properties. For cargo transport by
motors teams, binding defects also change the effective unbinding rate of the
cargo particles and are expected to have a stronger effect.Comment: 20 pages, latex, 7 figures, 1 tabl
Force and Motion Generation of Molecular Motors: A Generic Description
We review the properties of biological motor proteins which move along linear
filaments that are polar and periodic. The physics of the operation of such
motors can be described by simple stochastic models which are coupled to a
chemical reaction. We analyze the essential features of force and motion
generation and discuss the general properties of single motors in the framework
of two-state models. Systems which contain large numbers of motors such as
muscles and flagella motivate the study of many interacting motors within the
framework of simple models. In this case, collective effects can lead to new
types of behaviors such as dynamic instabilities of the steady states and
oscillatory motion.Comment: 29 pages, 9 figure
Imbalance of Hsp70 family variants fosters tau accumulation
Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154405/1/fsb2027004018.pd
Macrocyclic Ī²-Sheet Peptides That Inhibit the Aggregation of a Tau-Protein-Derived Hexapeptide
This paper describes studies of a series of macrocyclic Ī²-sheet peptides 1 that inhibit the aggregation of a tau-protein-derived peptide. The macrocyclic Ī²-sheet peptides comprise a pentapeptide "upper" strand, two Ī“-linked ornithine turn units, and a "lower" strand comprising two additional residues and the Ī²-sheet peptidomimetic template "Hao". The tau-derived peptide Ac-VQIVYK-NH(2) (AcPHF6) aggregates in solution through Ī²-sheet interactions to form straight and twisted filaments similar to those formed by tau protein in Alzheimer's neurofibrillary tangles. Macrocycles 1 containing the pentapeptide VQIVY in the "upper" strand delay and suppress the onset of aggregation of the AcPHF6 peptide. Inhibition is particularly pronounced in macrocycles 1a, 1d, and 1f, in which the two residues in the "lower" strand provide a pattern of hydrophobicity and hydrophilicity that matches that of the pentapeptide "upper" strand. Inhibition varies strongly with the concentration of these macrocycles, suggesting that it is cooperative. Macrocycle 1b containing the pentapeptide QIVYK shows little inhibition, suggesting the possibility of a preferred direction of growth of AcPHF6 Ī²-sheets. On the basis of these studies, a model is proposed in which the AcPHF6 amyloid grows as a layered pair of Ī²-sheets and in which growth is blocked by a pair of macrocycles that cap the growing paired hydrogen-bonding edges. This model provides a provocative and appealing target for future inhibitor design
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