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Taly, A.

Publications and source records attributed to Taly, A..

15 recordsLinked to original sources

Can SMILES be fragmented into a concatenable ordered sequence of retrosynthetically interesting string block ?

Molecules generated by deep learning models are often difficult to synthesize. Their synthetic accessibility can be improved with automated retrosynthetic analysis, which allows for identifying synthons. However, synthons in a SMILES can be scattered throughout the string depending on the path taken through the molecular graph used to generate the SMILES. We tested whether the ensemble of possible SMILES for a molecule can be used to generate a concatenable ordered sequence of string fragments (blocks) from SMILES that match potential synthons obtained through automated retrosynthetic analysis. We found that exhaustively sampling the SMILES space of a molecule improves the coverage of retrosynthetic breaks. We achieved full coverage of retrosynthetic bonds in string form for 85\% of the 1.9 million molecules in the MOSES dataset. Doing so allowed us to test our block SMILES in an unconditional de novo drug design test case with MolGPT and Monte Carlo Tree Search (MCTS). We found that using blocks as an LLM's token did degrade MolGPT performance due to the curse of dimensionality. However, using the SMILES selected by our blocking algorithm with the default SMILES tokenizer improved the reproduction of physico-chemical properties of samples and also improved uniqueness, novelty, and validity. The MCTS outperforms our MolGPT models in terms of validity and novelty. However, samples generated by the MCTS had physico-chemical properties that were further away from the MOSES baseline than the samples produced by molGPT, with an improved distribution of quantitative estimation of drug-likeness (QED).

bioinformatics↗

Deep-learning predictions of biomolecular structures : persistent limitations and new horizons extended by explicit ion addition

The advent of deep learning-driven tools such as AlphaFold has revolutionized the prediction of biomolecular structures, offering unprecedented accuracy and accessibility for proteins, RNA, and their complexes. While these tools have demonstrated remarkable success in benchmarking competitions and enabled experimentalists to generate models with ease, their widespread use has also highlighted persistent challenges. These include difficulties in assessing model confidence, limitations in predicting transmembrane domains, nucleic acids, conformational diversity, and interactions with ions or ligands, as well as the tendency to misfold intrinsically disordered regions (IDRs). In this perspective, we critically evaluate the strengths and limitations of current AI-based structure prediction tools through illustrative examples with a particular emphasis on the impact of explicit ion modelling. We notably report how the explicit addition of a few potassium cations to the prediction of IDRs or G-quadruplexes can trigger massive conformational switches compared to "dry" predictions. On this basis, we suggest modelling sequences both "dry" and in the presence of explicit potassium cations as a simple, practical way to sample alternative conformations and to expose disordered regions that current predictors tend to over-fold. We discuss the importance of reporting confidence metrics in publications to avoid overinterpretation. Furthermore, we address the unique challenges of RNA structure prediction, where data scarcity and structural complexity limit the performance of both classical and deep learning methods. Our analysis underscores the need for continued methodological advancements, integration of complementary computational tools, and expansion of high-quality experimental datasets. TOC Graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=111 SRC="FIGDIR/small/740587v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@e950e9org.highwire.dtl.DTLVardef@1bf22b3org.highwire.dtl.DTLVardef@17f5229org.highwire.dtl.DTLVardef@1eb2e9f_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

Full-Length Structural Modeling of Mitofusins with AlphaFold Reveals a Novel Cross-Type Dimerization and Insights into Oligomerization

Mitochondrial dynamics, involving fission and fusion, are critical for the maintenance, function, distribution, and inheritance of mitochondria, allowing their morphology to adapt to the cells physiological needs. Mitofusins, large GTPase transmembrane proteins, play a role in this process by driving the tethering and fusion of mitochondrial outer membranes. Dysfunction in mitofusins has been associated with neurodegenerative diseases such as Parkinsons, Alzheimers, Huntingtons, and Charcot-Marie-Tooth type 2A, as well as various cancers, where their dysregulated expression influences cell proliferation, invasion, and chemotherapy resistance. Despite their importance, the precise molecular mechanisms underlying mitofusin-mediated fusion remain unclear and require further structural elucidation. In fact, no complete high-resolution structures exist for mitofusins, including their yeast homolog, Fzo1. Here, we generated and analyzed full-length structural models of mitofusins using AlphaFold. Monomeric, dimeric, and tetrameric assemblies were produced, including complexes with fusion partners Ugo1 and SLC25A46. While AlphaFold predicted limited conformational diversity for isolated monomers, structural variability emerged for predicted homo- and hetero-oligomers. Notably, our models reveal a previously undescribed cross-type dimerization mode involving interactions between heptad repeat domains, not reported in current experimental structures. Comparison with recently resolved experimental data further supports the structural relevance of this interface. These full-length models allowed us to propose a new hypothetical mechanism of outer mitochondrial membrane fusion.

bioinformatics↗

Statistical Molecular Interaction Fields: A Fast andInformative Tool for Characterizing RNA and ProteinBinding Pockets

Developing a physical understanding of the interactions between a macro-molecular target and its ligands is a crucial step in structure-based drug design. Although many tools exist to characterize protein-binding pockets in silico, this is not yet the case for RNA, which has only recently been recognized as a suitable target for small ligands. Molecular Interaction Fields (MIF) are a useful tool to characterize the interactions of a given binding pocket. However, classical MIFs heavily rely on the use of probes, which makes their calculation accurate but very specific to the binding partners in question. We develop here a simple version of MIF, that we call Statistical Molecular Interaction Fields (SMIF), based on functional forms inspired by coarse-grained models and parametrized based on PDB structures and previous statistical analysis of the main form of interactions typical of macromolecules, namely hydrogen bonding, stacking, and hydrophobic interactions. We show that these fields, despite their simplicity, are very informative and overall in agreement with pharmacophoric models. Thanks to a carefully optimized code, our calculations are fast and can be performed in bulk on a large set of binding pockets or even on a full macromolecule. As shown in a few representative examples, the latter possibility opens the way to the analysis of systems as large as 20000 to 80000 atoms in relation to the surrounding environment, i.e., a lipidic membrane, a small ligand, or another macromolecular partner, allowing for a detailed visualization of the possible interactions. The complete software and its documentation are available here: https://smiffer.mol3d.tech/

biophysics↗

Menger_Curvature : a MDAKit implementation to decipher the dynamics, curvatures and flexibilities of polymeric backbones at the residue level

Characterizing the dynamics of the backbone of flexible polymers such as Intrinsically Disordered Regions and Proteins (IDRs and IDPs) has proven to be a significant challenge in molecular dynamics (MD) simulations due to their high conformational variability. The widely-used mobility metric Root-Mean-Squared Fluctuations (RMSF) is powerless to provide information for highly flexible systems, as defining a relevant reference structure is often not possible. We previously introduced a new flexibility metric to remedy this gap : the Local Flexibilities (LFs), derived (alongside the Local Curvatures (LCs)) from the Proteic Menger Curvatures (PMCs). Here we present a numba accelerated implementation for any polymer of the calculation of Menger Curvatures as a MDAKit from the widely-used MDAnalysis package. We perform a benchmark with the RMSF and another flexibility metric derived from Proteic Blocks (PBs), the Equivalent Number of PBs (Neq), and show that the PMCs are an order of magnitude faster to compute on a modern CPU chip. We applied all 3 flexibility metrics to a {beta}III-tubulin monomer as an example, as tubulins are known to possess the entire range of proteic elements, from -helix and {beta}-sheets to a flexible loop and a disordered C-terminal tail (CTT). RMSF, LFs and Neq all succeed in identifying the flexible loops and the CTT, although the RMSF requires a system-specific alignment to do so. Finally, we expose different applications of PMCs, LCs and LFs ranging from mechanism characterization to NMR T2 predictions. We believe that Menger curvatures will prove to be a valuable metric to study protein dynamics and polymers in general. The MDAKit package Menger_Curvature is readily available at https://github.com/EtienneReboul/menger_curvature

bioinformatics↗

Improving the reliability of molecular string representations for generative chemistry

Generative modeling for chemistry has advanced rapidly in recent years, but this surge in popularity raises a foundational question: which molecular representation is best suited for modern machine learning models? Despite not being designed for generative tasks, SMILES remain the most commonly used string-based representation. However, while SMILES follow strict syntactic rules, grammatically correct SMILES strings do not always correspond to valid molecules. SELFIES were introduced as an alternative that addresses this limitation by ensuring that every string of SELFIES tokens represents to a valid molecule. In this study, we comprehensively evaluate the limitations of both SMILES and SELFIES as representations for generative models. We define two key criteria for robust molecular generation: viability, generated strings represent novel, unique molecules with correct valence, and fidelity, the distribution of physicochemical properties from sampled molecules resembles that of the training data. We find that approximately one-fifth of molecules generated using canonical SMILES are invalid, failing the viability criterion. In contrast, all SELFIES-generated molecules are viable, but they deviate significantly from the training distribution, indicating low fidelity. To address these limitations, we develop data augmentation procedures for both representations. While simplifying the SELFIES grammar yields only modest gains in fidelity, our stochastic augmentation method for SMILES, ClearSMILES, significantly improves both viability and fidelity. ClearSMILES simplifies syntax by reducing the vocabulary size and explicitly encoding aromaticity via Kekule SMILES, making it easier string representations for models to process. Using ClearSMILES, the rate of invalid samples decreases by an order of magnitude, from 20% to 2.2%, and fidelity to the training distribution is also moderately improved. Generative chemistry has seen rapid development recently. However, models based on string representations of molecules still rely largely on SMILES1 that have not been developed for this context and SELFIES 2 who were introduced to reduce those problems. The goal of this study is to first analyze the difficulty encountered by a small generative model when using SMILES and SELFIES. Our study found that SELFIES and canonical SMILES 3 are not fully reliable representations for a small generative model, i.e. do not ensure concurrently the viability and fidelity of samples. Viable samples represent novel, unique molecules with correct valence, while fidelity is efficient distribution learning of key physico-chemical properties. 4 In fact, 20% of the samples generated using canonical SMILES input representation do not correspond to valid molecules. In contrast, samples generated using SELFIES were all viable but where not able to reproduce as well the distribution of physico-chemical properties as SMILES. As a mitigation strategy for the previously identified problems, we have developed data augmentation procedures for both SELFIES and SMILES. Simplifying the complex syntax of SELFIES yielded only marginal improvements in string stability and overall fidelity to the training set. For SMILES, we developed a stochastic data augmentation procedure called ClearSMILES, which reduces the vocabulary size needed to represent a SMILES dataset, explicitly represents aromaticity via Kekule SMILES, 3 and reduces the effort required by deep learning models to process SMILES. ClearSMILES reduced the rate of invalid samples by an order of magnitude, from 20% to 2.2%, and improved the fidelity of samples to the training set.

bioinformatics↗

Molecular modeling of Proteinase-Activated Receptor 1 in complex with Thrombin Receptor Activator Peptide 6.

The protease-activated receptor 1 (PAR1) and its activator thrombin re-ceptor activator peptide 6 (TRAP6) play crucial roles in various physiologi-cal and pathological processes, including hemostasis, thrombosis, and cancer progression. Although the interaction between PAR1 and TRAP6 has been heavily studied using experimental technique such as mutagenesis, structural data remains scarce due to the technical hardship of studying membrane pro-teins such as PAR1. In this study, we employed an integrative modeling approach to elucidate the structure of the PAR1-TRAP6 complex. Leveraging state-of-the-art AI-based protein modeling tools, including AlphaFold2 and ESMFOLD, we in-tegrated HADDOCK, a physics-based method to refine predictions. Overall, the predicted structures are in good agreement with the experimental data available in the literature. Our model unveiled a new T-shaped pi-stacking interaction between TRAP6s F2 and PAR1s Y360. The integrative modeling approach combining the predictions of the deep learning model with a physics-based method proves to be an interesting strat-egy for solving challenging membrane protein structures with high confidence. Our model of the PAR1-TRAP6 complex will be an interesting starting point for further investigation of the activation of PAR1 by TRAP6.

bioinformatics↗

Combining computational biology and experimental knowledge to draw a structural profile of the active membrane-assembled NADPH oxidase complex

The phagocyte NADPH oxidase complex (NOX2) is activated by various intracellular stimuli, resulting in the generation of reactive oxygen species, which are responsible for many physiological processes, including innate immune defense. Structures of the cytochrome b558, membrane catalytic core of the complex, in their resting state have been solved experimentally. Unfortunately, very little is known about the structure of the active complex, as it involves the assembly of modular proteins and disordered regions, that have been shown to be essential to the function of the enzyme. Here, we modelled the supra-structure of the active complex using Alphafold2 and taking into account experimental data as well as conformational analyses by SRCD. AF2 ability to generate complexes enabled us to visualize ways in which subunits could assemble together, and revealed the implications of intrinsically disordered regions in protein-protein and membrane-protein interactions. Additional Molecular Dynamics simulations of the membrane embedded complex bring further insight regarding the system dynamics. Altogether, our work provides evidence for a model of the structure of the active complex where disordered regions can enable multi-subunit assembly.

biochemistry↗

Plasmodesmata act as unconventional membrane contact sites regulating inter-cellular molecular exchange in plants

Membrane contact sites (MCS) are fundamental for intracellular communication, but their role in intercellular communication remains unexplored. We show that in plants, plasmodesmata communication bridges function as atypical endoplasmic reticulum (ER)-plasma membrane (PM) tubular MCS, operating at cell-cell interfaces. Similar to other MCS, ER-PM apposition is controlled by a protein-lipid tethering complex, but uniquely, this serves intercellular communication. Combining high-resolution microscopy, molecular dynamics, pharmacological and genetic approaches, we show that cell-cell trafficking is modulated through the combined action of Multiple C2 domains and transmembrane domain proteins (MCTP) 3, 4, and 6 ER-PM tethers, and phosphatidylinositol-4-phosphate (PI4P) lipid. Graded PI4P amounts regulate MCTP docking to the PM, their plasmodesmata localization and cell-cell permeability. SAC7, an ER-localized PI4P-phosphatase, regulates MCTP4 accumulation at plasmodesmata and modulates cell-cell trafficking capacity in a cell-type specific manner. Our findings expand MCSs functions in information transmission, from intracellular to intercellular cellular activities. In briefPlant intercellular communication is regulated via tubular membrane contact through PI4P binding-ER-PM tether MCTP proteins HighlightsO_LIPlasmodesmata are unconventional ER/PM tubular contact sites located at cell-cell interface C_LIO_LIPlasmodesmata operate as control valves, modulating ER-PM contacts to regulate transport C_LIO_LIMCTP3, MCTP4, MCTP6 and PI4P tethering elements act as valve regulators C_LIO_LISAC7 PI4P phosphatase controls plasmodesmata MCS in a cell-type-specific manner C_LI

plant biology↗

Plant plasmodesmata bridges form through ER-driven incomplete cytokinesis

Diverging from conventional cell division models, plant cells undergo incomplete division to generate plasmodesmata communication bridges between daughter cells. While fundamental for plant multicellularity, the molecular events leading to bridge stabilization, as opposed to severing, remain unknown. Using electron tomography, we mapped the transition from cell plate fenestrae to plasmodesmata. We show that the ER connects daughter cells across fenestrae, and as the cell plate matures, fenestrae contract, causing the PM to mold around constricted ER tubes. The ERs presence prevents fenestrae fusion, forming plasmodesmata, while its absence results in closure. The ER-PM tethers MCTP3, 4, and 6 further stabilize nascent plasmodesmata during fenestrae contraction. Genetic deletion in Arabidopsis reduces plasmodesmata formation. Our findings reveal how plants undergo incomplete division to promote intercellular communication. One-Sentence SummaryThe ER is important for stabilizing nascent plasmodesmata, a process integral to incomplete cytokinesis in plants.

plant biology↗

Lys716 in the transmembrane domain of yeast mitofusin Fzo1 modulates anchoring and fusion

Outer mitochondrial membrane (OMM) fusion is an important process for the cell and organism survival, as its dysfunction is linked to neurodegenerative diseases and cancer. The OMM fusion is mediated by members of the dynamin-related protein (DRP) family, named mitofusins. The exact mechanism by which the mitofusins contribute to these diseases, as well as the exact molecular fusion mechanism mediated by mitofusin, remains elusive. We have performed extensive multiscale molecular dynamics simulations using both coarse-grained and all-atom approaches to predict the dimerization of two transmembrane domain (TM) helices of the yeast mitofusin Fzo1. We identify specific residues, such as Lys716, that can modulate dimer stability. Comparison with a previous computational model reveals remarkable differences in helix crossing angles and interfacial contacts. Overall, however, the TM1-TM2 interface appears to be stable in the Martini and CHARMM force fields. Replica-exchange simulations further tune a detailed atomistic model, as confirmed by a remarkable agreement with an independent prediction of the Fzo1-Ugo1 complex by AlphaFold2. Functional implications, including a possible role of Lys716 that could affect membrane interactions during fusion, are suggested and consistent with experiments monitoring mitochondrial respiration of selected Fzo1 mutants.

bioinformatics↗

Deep Learning-Based Prediction of A. thaliana's MCTP4 Structure and Exploration of Transmembrane Dynamics using Coarse-Grained Molecular Dynamics Simulations

Multiple C2 Domains and Transmembrane region Proteins (MCTPs) in plants have been identified as important functional and structural components of plasmodesmata cytoplasmic bridges, which are vital for cell-cell communication. MCTPs are endoplasmic reticulum (ER)-associated proteins which contain three to four C2 domains and two transmembrane regions. In this study, we created structural models of Arabidopsis MCTP4 ER-anchor transmembrane region (TMR) domain using several prediction methods based on deep learning (DL). This region, critical for driving ER association, presents a complex domain organization and remains largely unknown. Our study demonstrates that using a single deep-learning method to predict the structure of membrane proteins can be challenging. Our deep learning models presented three different conformations for the MCTP4 structure, provided by different deep learning methods, indicating the potential complexity of the proteins conformational landscape. For the first time, we used simulations to explore the behaviour of the TMR of MCTPs within the lipid bilayer. We found that the TMR of MCTP4 is not rigid, but can adopt various conformations including some not identified by deep learning tools. These findings underscore the complexity of predicting protein structures. We learned that combining different methods, such as deep learning and simulations, enhances our understanding of complex proteins.

bioinformatics↗

Exportin 1-mediated nuclear/cytoplasmic trafficking controls drug sensitivity of classical Hodgkin lymphoma

Exportin 1 (XPO1) is the main nuclear export receptor that controls the subcellular trafficking and the functions of major regulatory proteins. XPO1 is overexpressed in various cancers and small inhibitors of nuclear export (SINEs) have been developed to inhibit XPO1. In primary mediastinal B-cell lymphoma (PMBL) and classical Hodgkin lymphoma (cHL), the XPO1 gene may be mutated on one nucleotide and encodes the mutant XPO1E571K. To understand the impact of mutation on protein function, we studied the response of PMBL and cHL cells to selinexor, a SINE, and ibrutinib, an inhibitor of Bruton tyrosine kinase. XPO1 mutation renders lymphoma cells more sensitive to selinexor due to a faster degradation of mutant XPO1 compared to the wild-type. We further showed that a mistrafficking of p65 (RELA) and p52 (NF{kappa}B2) transcription factors between the nuclear and cytoplasmic compartments accounts for the response towards ibrutinib. XPO1 mutation may be envisaged as a biomarker of the response of PMBL and cHL cells and other B-cell hemopathies, to SINEs and drugs that target even indirectly the NF{kappa}B signaling pathway.

cancer biology↗

Mutational analysis to explore long-range allosteric coupling and decoupling in a pentameric channel receptor

Pentameric ligand-gated ion channels (pLGICs) mediate chemical signaling through a succession of allosteric transitions that are yet not completely understood. On the prototypic bacterial channel GLIC, we explored the conformational landscape of the protein during pH-gating. To this aim, we introduced a series of allosteric mutations, and characterized the protein conformation over a broad pH range. We combined electrophysiological recordings, fluorescence quenching experiments monitoring key quaternary reorganizations, and simulations by normal mode analysis. Moderate loss-of-function mutations and the allosteric modulator propofol displace allosteric equilibria involved in pre-activation and pore opening processes, highlighting long-range allosteric coupling between distant regions of the protein. In contrast, total loss-of-function mutations stabilize the protein in unique intermediate conformations where motions are decoupled. Altogether, our data show that the protein can access a wide conformational landscape, raising the possibility of multiple conformational pathways during gating.Competing Interest StatementThe authors have declared no competing interest.View Full Text

biophysics↗

Probing the ionotropic activity of the orphan glutamate delta 2 receptor with genetically-engineered photopharmacology.

Glutamate delta (GluD) receptors belong to the ionotropic glutamate receptor family, yet whether they actually form functional and physiologically-relevant ion channels in neurons remains a debated question. Here we used a chemo-genetic approach to engineer specific and photo-reversible pharmacology in the orphan GluD2 receptor. We incorporated a cysteine mutation in the cavity located above the putative ion channel pore, for site-specific conjugation with a photoswitchable ligand. We first showed that, in the constitutively-open GluD2 Lurcher mutant, current could be rapidly and reversibly decreased with light. We then transposed the cysteine mutation to the native receptor, to demonstrate with absolute pharmacological specificity that metabotropic glutamate receptor signaling opens the GluD2 ion channel in heterologous expression system. Our results assess the functional relevance of GluD2 ion channel and introduce an optogenetic tool that will provide a novel and powerful means for probing GluD2 ionotropic contribution to neuronal physiology.

neuroscience↗