Ajay S. Thatte

Bio: Ajay S. Thatte is an academic researcher from University of Texas at Austin. The author has contributed to research in topics: Membrane curvature & Signal transducing adaptor protein. The author has an hindex of 3, co-authored 4 publications receiving 49 citations.

Molecular Mechanisms of Membrane Curvature Sensing by a Disordered Protein

Abstract: The ability of proteins to sense membrane curvature is essential for the initiation and assembly of curved membrane structures. Established mechanisms of curvature sensing rely on proteins with spe...

Structured and intrinsically disordered domains within Amphiphysin1 work together to sense and drive membrane curvature.

Abstract: Cellular membranes undergo remodeling during many cellular processes including endocytosis, cytoskeletal protrusion, and organelle biogenesis. During these events, specialized proteins sense and amplify fluctuations in membrane curvature to create stably curved architectures. Amphiphysin1 is a multi-domain protein containing an N-terminal crescent-shaped BAR (Bin/Amphiphysin/Rvs) domain and a C-terminal domain that is largely disordered. When studied in isolation, the BAR domain of Amphiphysin1 senses membrane curvature and generates membrane tubules. However, the disordered domain has been largely overlooked in these studies. Interestingly, our recent work has demonstrated that the disordered domain is capable of substantially amplifying the membrane remodeling ability of the BAR domain. However, the physical mechanisms responsible for these effects are presently unclear. Here we elucidated the functional role of the disordered domain by gradually truncating it, creating a family of mutant proteins, each of which contained the BAR domain and a fraction of the disordered domain. Using quantitative fluorescence and electron microscopy, our results indicate that the disordered domain contributes to membrane remodeling by making it more difficult for the protein to bind to and assemble on flat membrane surfaces. Specifically, we found that the disordered domain began to significantly impact membrane remodeling when its projected area exceeded that of the BAR domain. Once this threshold was crossed, steric interactions with the membrane surface and with neighboring disordered domains gave rise to increased curvature sensing and membrane vesiculation, respectively. These findings provide insight into the synergy between structured and disordered domains, each of which play important biophysical roles in membrane remodeling.

Clathrin senses membrane curvature

Abstract: The ability of proteins to sense membrane curvature is essential to diverse membrane remodeling processes including clathrin-mediated endocytosis. Multiple adaptor proteins within the clathrin pathway have been shown to assemble together at curved membrane sites, leading to local recruitment of the clathrin coat. Because clathrin does not bind to the membrane directly, it has remained unclear whether clathrin plays an active role in sensing curvature or is passively recruited by its adaptor proteins. Using a synthetic tag to assemble clathrin directly on membrane surfaces, here we show that clathrin is a strong sensor of membrane curvature, comparable to previously studied adaptor proteins. Interestingly, this sensitivity arises from clathrin assembly, rather than from the properties of unassembled triskelia, suggesting that triskelia have preferred angles of interaction, as predicted by earlier structural data. Further, when clathrin is recruited by adaptors, its curvature sensitivity is amplified by two to ten-fold, such that the resulting protein complex is up to 100 times more likely to assemble on a highly curved surface, compared to a flatter one. This exquisite sensitivity points to a synergistic relationship between the coat and its adaptor proteins, which enables clathrin to pinpoint sites of high membrane curvature, an essential step in ensuring robust membrane traffic. More broadly, these findings suggest that protein networks, rather than individual protein domains, are likely the critical drivers of membrane curvature sensing.

Ionizable Lipid Nanoparticles for In Vivo mRNA Delivery to the Placenta during Pregnancy.

Abstract: Ionizable lipid nanoparticles (LNPs) are the most clinically advanced nonviral platform for mRNA delivery. While they have been explored for applications including vaccines and gene editing, LNPs have not been investigated for placental insufficiency during pregnancy. Placental insufficiency is caused by inadequate blood flow in the placenta, which results in increased maternal blood pressure and restricted fetal growth. Therefore, improving vasodilation in the placenta can benefit both maternal and fetal health. Here, we engineered ionizable LNPs for mRNA delivery to the placenta with applications in mediating placental vasodilation. We designed a library of ionizable lipids to formulate LNPs for mRNA delivery to placental cells and identified a lead LNP that enables in vivo mRNA delivery to trophoblasts, endothelial cells, and immune cells in the placenta. Delivery of this top LNP formulation encapsulated with VEGF-A mRNA engendered placental vasodilation, demonstrating the potential of mRNA LNPs for protein replacement therapy during pregnancy to treat placental disorders.

Physical Biology of the Cell

Abstract: Physical Biology of the Cell, 2nd Edition, is a textbook that focuses on the application of physical principles to understanding biological systems. The subject matter of the text is organized according to common physical principles that govern biological processes rather than in relation to the biological processes themselves, as is common for most biology and cell biology textbooks. Topics covered in the book span a broad range of interests, including electrostatics, molecular interactions, molecular motors and the cytoskeleton, and membranes. Each chapter features color figures, derived equations with relevant examples, and problem sets at the chapter’s conclusion. The problem sets at the end of the chapters are expanded from the first edition. Further, the second edition includes two new chapters, one on light and pattern formation, and another on the use of computation in exploring biological problems. Additional student and instructor resources are also available online. The primary audience for the textbook could include advanced undergraduate students or first-year graduate students. While the textbook may be best suited for a biophysics course, it could also be used as a primary or supplementary text for teaching cellular and molecular biology. As a teaching tool for cellular and molecular biology, the many examples featured throughout the text could easily be employed to assist students in learning the principles of how a cellular or molecular system functions. A basic level of mathematical proficiency would be required of the student. While this textbook could be an excellent resource for many courses, there are several topics commonly covered in biochemistry classes, such the glycolytic pathway, that are featured in the book but in different contexts than many widely used biochemistry texts. For a biophysics course that is heavily focused on techniques, individual references that discuss the specific techniques in detail would be more suitable than this book. Of course, any instructor seeking to use this textbook should be aware of its content before making a selection. This textbook is an excellent resource, both for a research scientist and for a teacher. The authors do a superb job of selecting the material for each chapter and explaining the material with equations and narrative in an easily digestible manner. Readers who enjoy this book may also enjoy Molecular Driving Forces by Dill and Bromberg, which gives excellent treatment of similar concepts.

Membrane-Binding Peptides for Extracellular Vesicles On-Chip Analysis

Abstract: Small extracellular vesicles (sEVs) present fairly distinctive lipid membrane features in the extracellular environment. These include high curvature, lipid-packing defects and a relative abundance in lipids such as phosphatidylserine and ceramide. sEV membrane could be then considered as a "universal" marker, alternative or complementary to traditional, characteristic, surface-associated proteins. Here, we introduce the use of membrane-sensing peptides as new, highly efficient ligands to directly integrate sEV capturing and analysis on a microarray platform. Samples were analysed by label-free, single-particle counting and sizing, and by fluorescence co-localisation immune staining with fluorescent anti-CD9/anti-CD63/anti-CD81 antibodies. Peptides performed as selective yet general sEV baits and showed a binding capacity higher than anti-tetraspanins antibodies. Insights into surface chemistry for optimal peptide performances are also discussed, as capturing efficiency is strictly bound to probes surface orientation effects. We anticipate that this new class of ligands, also due to the versatility and limited costs of synthetic peptides, may greatly enrich the molecular toolbox for EV analysis.

Intrinsically disordered proteins and membranes: a marriage of convenience for cell signalling?

Abstract: The structure-function paradigm has guided investigations into the molecules involved in cellular signalling for decades. The peripheries of this paradigm, however, start to unravel when considering the co-operation between proteins and the membrane in signalling processes. Intrinsically disordered regions hold distinct advantages over folded domains in terms of their binding promiscuity, sensitivity to their particular environment and their ease of modulation through post-translational modifications. Low sequence complexity and bias towards charged residues are also favourable for the multivalent electrostatic interactions that occur at the surfaces of lipid bilayers. This review looks at the principles behind the successful marriage between protein disorder and membranes in addition to the role of this partnership in modifying and regulating signalling in cellular processes. The HVR (hypervariable region) of small GTPases is highlighted as a well-studied example of the nuanced role a short intrinsically disordered region can play in the fine-tuning of signalling pathways.