Quantitative biology of endocytosis / Julien Berro, Michael M. Lacy

By: Contributor(s): Resource type: Ressourcentyp: Buch (Online)Book (Online)Language: English Series: Colloquium series on quantitive cell biology ; # 4Publisher: [San Rafael, California] : Morgan & Claypool, 2018Description: 1 Online-Ressource (1 PDF (xi, 73 pages)))Subject(s): Additional physical formats: 9781615047864 | 9781615047840 | 1615047859. | 9781615047857. | Erscheint auch als: 9781615047840 Druck-AusgabeDDC classification:
  • 571.655
LOC classification:
  • QH634
NLM classification:
  • QU 375
Online resources: Summary: 2. Collecting quantitative data -- 2.1 Using fluorescence microscopy to obtain quantitative data about endocytosis -- Overview of fluorescence microscopes that can be used for quantitative analysis of endocytosis -- Comparison of different fluorescence microscopy systems -- General considerations about fluorescent tags -- 2.2 Practical considerations for counting molecules using a fluorescence microscope -- Tag a protein of interest at its genomic locus -- Limit photobleaching -- Limit uneven illumination of the field of view -- Ensure the microscopy system is used in conditions where the signal is linear -- Calibrate the microscope using flexible settings to avoid calibrating too often -- Best practices to limit sources of variability -- 2.3 Electron microscopy and correlative light and electron microscopy (CLEM) -- 2.4 Super-resolution microscopy --Summary: 4. Using quantitative microscopy data to infer the molecular mechanisms of endocytosis -- 4.1 How are endocytic proteins organized? -- The protein copy numbers and their stoichiometries constrain possible molecular organizations and mechanisms -- High-resolution track alignment can infer the organization of endocytic proteins -- The organization of the actin meshwork around the endocytic vesicle can be inferred from the vesicle's movements -- 4.2 What are the molecular mechanisms of actin dynamics during endocytosis? -- Inferring mechanisms by comparing numbers and stoichiometries -- Mathematical modeling constrained by quantitative data allows one to test different mechanisms and make testable predictions -- 4.3 What are the mechanical properties of the membrane and the forces involved in membrane deformations? -- Modeling of membrane shapes can elucidate mechanical properties and forces involved during CME -- Simulations to test possible force distributions to elongate the CCP and propose new mechanisms --Summary: 5. Perspectives and future of quantitative biology of endocytosis -- 5.1 Open questions in CME -- 5.2 Quantities that are not (yet?) measurable -- 5.3 Advances in microscopy techniques -- 5.4 Advances in computational tools -- 5.5 Applications to other systems --Summary: Clathrin-mediated endocytosis (CME) is a ubiquitous internalization process in eukaryotic cells. It consists of the formation of an approximately 50-nm diameter vesicle out of a flat membrane. Genetics, biochemistry, and microscopy experiments performed in the last four decades have been instrumental to discover and characterize major endocytic proteins in yeast and mammals. However, due to the highly dynamic nature of the endocytic assembly and its small size, many questions remain unresolved: how are endocytic proteins organized spatially and dynamically? How are forces produced and how are their directions controlled? How do the biochemical activities of endocytic proteins and the membrane shape and mechanics regulate each other? These questions are virtually impossible to visualize or measure directly with conventional approaches but thanks to new quantitative biology methods, it is now possible to infer the mechanisms of endocytosis in exquisite detail. This book introduces quantitative microscopy and mathematical modeling approaches that have been used to count the copy number of endocytic proteins, infer their localization with nanometer precision, and infer molecular and physical mechanisms that are involved in the robust formation of endocytic vesiclesPPN: PPN: 1032028262Package identifier: Produktsigel: ZDB-4-NLEBK
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