The current paradigm in the amyloid hypothesis brands small beta-amyloid (Abeta) oligomers as the toxic species in Alzheimer's disease (AD). These oligomers are fibril-like;contain beta-sheet structure, and present exposed hydrophobic surface. Oligomers with this motif are capable of penetrating the cell membrane, gathering to form toxic ion channels. Current agents suppressing precursor Abeta cleavage have only met partial success;and to date, those targeting the peptides and their assemblies in the aqueous environment of the extracellular space largely fail in clinical trials. One possible reason is failure to reach membrane-embedded targets of disease-'infected'cells. We pointed to the need to account for the lipid environment when aiming to prevent the formation of toxic channels, and propose a combination therapy to target the species spectrum. Amyloid-beta (Abeta) oligomers destabilize cellular ionic homeostasis, mediating Alzheimer's disease (AD). It is still unclear whether the mechanism (i) is mediated by cell surface receptors;(ii) is direct, with Abeta oligomers interacting with membrane lipids;or (iii) both mechanisms take place. Recent studies indicate that Abeta oligomers may act by either of the last two. Little is known about the oligomers'structures and how they spontaneously insert into the membrane. Using explicit solvent molecular dynamics (MD) simulations, we show that fibril-like Abeta(17-42) (p3) oligomer is capable of penetrating the membrane. Insertion is similar to that observed for protegrin-1 (PG-1), a cytolytic beta-sheet-rich antimicrobial peptide (AMP). Both Abeta and PG-1 favor the amphipathic interface of the lipid bilayer in the early stage of interaction with the membrane. U-shaped Abeta oligomers are observed in solution and in the membrane, suggesting that the preformed seeds can be shared by amyloid fibrils in the growth phase and membrane toxicity. Here we provide sequential events in possible Abeta oligomer membrane-insertion pathways. We speculate that for the U-shaped motif, a trimer is the minimal oligomer size to insert effectively. We propose that monomers and dimers may insert in (apparently on-pathway) aggregation-intermediate beta-hairpin state, and may (or may not) convert to a U-shape in the bilayer. Together with earlier observations, our results point to a non-specific, broadly heterogeneous landscape of membrane-inserting oligomer conformations, pathways, and membrane-mediated toxicity of beta-rich oligomers. Tau pathology in Alzheimer's disease is intimately linked to the deposition of proteinacious filaments, which akin to infectious prions, have been proposed to spread via seeded conversion. Here we use double electron-electron resonance (DEER) spectroscopy in combination with extensive computational analysis to show that filaments of three- (3R) and four-repeat (4R) tau are conformationally distinct. Distance measurements between spin labels in the third repeat, reveal tau amyloid filaments as ensembles of known beta-strand-turn-beta-strand U-turn motifs. Whereas filaments seeded with 3R tau are structurally homogeneous, filaments seeded with 4R tau are heterogeneous, composed of at least three distinct conformers. These findings establish a molecular basis for the seeding barrier between different tau isoforms and offer a new powerful approach for investigating the composition and dynamics of amyloid fibril ensembles. In Alzheimer's disease and frontotemporal dementias, the microtubule-associated protein Tau forms intracellular paired helical filaments. The filaments can form not only by the full-length human Tau protein, but also by the three repeated (K19) or four repeated (K18) Tau segments. However, of interest, experimentally, K19 can seed K18, but not vice versa. To obtain insight into the cross-seeding between K18 and K19 aggregates, here, K18 and K19 octamers with repeat 3 (R3) in U-shaped, L-shaped, and long straight line-shaped (SL-shape) conformations are assembled into different structures. The simulation results show that K18-8/K19-8 (K18 and K19 assemblies number 8) with R3 in an L shape and K18-9/K19-9 with R3 in an SL shape are highly populated and present the highest structural similarity among all simulated K18 and K19 octamers, suggesting that similar folding of K18/K19 may serve as structural core for the K18-K19 co-assembled heterogeneous filament. We demonstrate that formation of stable R2 and R3 conformations is the critical step for K18 aggregation, and R3 is critical for K19 fibrillization. The different core units in K18 and K19 may create a cross-seeding barrier for the K18 seed to trigger K19 fibril growth because R2 is not available for K19. Our study provides insights into cross-seeding involving heterogeneous structures. The polymorphic nature of protein aggregation could be magnified in the cross-seeding process. If the seeding conformations lead to too much divergence in the energy landscape, it could impede fibril formation. Such an effect could also contribute to the asymmetric barrier between K18 and K19. Currently, there are two types of drugs on the market: orthosteric, which bind at the active site;and allosteric, which bind elsewhere on the protein surface, and allosterically change the conformation of the protein binding site. In this perspective we argue that the different mechanisms through which the two drug types affect protein activity and their potential pitfalls call for different considerations in drug design. The key problem facing orthosteric drugs is side effects which can occur by drug binding to homologous proteins sharing a similar binding site. Hence, orthosteric drugs should have very high affinity to the target;this would allow a low dosage to selectively achieve the goal of target-only binding. By contrast, allosteric drugs work by shifting the free energy landscape. Their binding to the protein surface perturbs the protein surface atoms, and the perturbation propagates like waves, finally reaching the binding site. Effective drugs should have atoms in good contact with the 'right'protein atoms;that is, the contacts should elicit propagation waves optimally reaching the protein binding site target. While affinity is important, the design should consider the protein conformational ensemble and the preferred propagation states.

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National Cancer Institute Division of Basic Sciences
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Tsai, Chung-Jung; Nussinov, Ruth (2014) The free energy landscape in translational science: how can somatic mutations result in constitutive oncogenic activation? Phys Chem Chem Phys 16:6332-41
Ma, Buyong; Nussinov, Ruth (2014) Druggable orthosteric and allosteric hot spots to target protein-protein interactions. Curr Pharm Des 20:1293-301
Tsai, Chung-Jung; Nussinov, Ruth (2014) A unified view of "how allostery works". PLoS Comput Biol 10:e1003394
Nussinov, Ruth; Ma, Buyong; Tsai, Chung-Jung (2014) Multiple conformational selection and induced fit events take place in allosteric propagation. Biophys Chem 186:22-30
Nussinov, Ruth; Tsai, Chung-Jung (2014) Free energy diagrams for protein function. Chem Biol 21:311-8
Nussinov, Ruth; Tsai, Chung-Jung (2014) Unraveling structural mechanisms of allosteric drug action. Trends Pharmacol Sci 35:256-64
Qi, Ruxi; Luo, Yin; Ma, Buyong et al. (2014) Conformational distribution and ?-helix to ?-sheet transition of human amylin fragment dimer. Biomacromolecules 15:122-31
Meyer, Virginia; Dinkel, Paul D; Luo, Yin et al. (2014) Single mutations in tau modulate the populations of fibril conformers through seed selection. Angew Chem Int Ed Engl 53:1590-3
Jang, Hyunbum; Arce, Fernando Teran; Ramachandran, Srinivasan et al. (2014) Disordered amyloidogenic peptides may insert into the membrane and assemble into common cyclic structural motifs. Chem Soc Rev 43:6750-64
Liu, Jin; Nussinov, Ruth (2013) The role of allostery in the ubiquitin-proteasome system. Crit Rev Biochem Mol Biol 48:89-97

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