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Phosphoinositides

Methods and Protocols

  • Book
  • © 2021

Overview

Editors:
  1. Roberto J. Botelho

    1. Department of Chemistry and Biology and the Graduate Program in Molecular Science, Ryerson University, Toronto, Canada

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  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2251)

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Table of contents (17 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Simultaneous Detection of Phosphoinositide Lipids by Radioactive Metabolic Labeling

    • Noah Steinfeld, Sai Srinivas Panapakkam Giridharan, Emily J. Kauffman, Lois S. Weisman
    Pages 1-17

  3. Label-Free Quantification of Phosphoinositides in Drosophila by Mass Spectrometry

    • Avishek Ghosh, Padinjat Raghu
    Pages 19-37

  4. Profiling of Phosphoinositide Molecular Species in Resting or Activated Human or Mouse Platelets by a LC-MS Method

    • Gaëtan Chicanne, Justine Bertrand-Michel, Julien Viaud, Karim Hnia, Jonathan Clark, Bernard Payrastre
    Pages 39-53

  5. Quantification of Genetically Encoded Lipid Biosensors

    • Rachel C. Wills, Jonathan Pacheco, Gerald R. V. Hammond
    Pages 55-72

  6. Detection of Plasma Membrane Phosphoinositide Dynamics Using Genetically Encoded Fluorescent Protein Probes

    • Rebecca Cabral-Dias, Yasmin Awadeh, Roberto J. Botelho, Costin N. Antonescu
    Pages 73-89

  7. Imaging the Nanoscale Distribution of Phosphoinositides in the Cell Plasma Membrane with Single-Molecule Localization Super-Resolution Microscopy

    • Fan Fan, Chen Ji, Xuelin Lou
    Pages 91-104

  8. Induced Dimerization Tools to Deplete Specific Phosphatidylinositol Phosphates

    • Jonathan Pacheco, Rachel C. Wills, Gerald R. V. Hammond
    Pages 105-120

  9. A Real-Time Phosphatidylinositol 4-Phosphate 5-Kinase Assay Using Fluorescence Spectroscopy

    • Taki Nishimura
    Pages 121-132

  10. Assessing In Situ Phosphoinositide–Protein Interactions Through Fluorescence Proximity Ligation Assay in Cultured Cells

    • Mo Chen, Hudson T. Horn, Tianmu Wen, Vincent L. Cryns, Richard A. Anderson
    Pages 133-142

  11. Characterization of Protein–Phospholipid/Membrane Interactions Using a “Membrane-on-a-Chip” Microfluidic System

    • Calvin Yeager, Djoshkun Shengjuler, Simou Sun, Paul S. Cremer, Craig E. Cameron
    Pages 143-156

  12. Exploring Phosphoinositide Binding Using Native Mass Spectrometry

    • Julian Bender, Carla Schmidt
    Pages 157-175

  13. Liposome-Based Methods to Study Protein–Phosphoinositide Interaction

    • Mélanie Mansat, Mélanie Picot, Gaëtan Chicanne, Virginie Nahoum, Frédérique Gaits-Iacovoni, Bernard Payrastre et al.
    Pages 177-184

  14. Liposome-Based Methods to Study GTPase Activation by Phosphoinositides

    • Julien Viaud, Laurie Ceccato, Bernard Payrastre, Frédérique Gaits-Iacovoni
    Pages 185-194

  15. Liposome Co-sedimentation and Co-flotation Assays to Study Lipid–Protein Interactions

    • Yosuke Senju, Pekka Lappalainen, Hongxia Zhao
    Pages 195-204

  16. Characterization of PROPPIN–Phosphoinositide Binding by Stopped-Flow Fluorescence Spectroscopy

    • Ángel Pérez-Lara, Reinhard Jahn
    Pages 205-214

  17. Fluorescence Assays to Study Membrane Penetration of Proteins

    • Yosuke Senju, Hongxia Zhao
    Pages 215-223

  18. Fluorogenic XY-69 in Lipid Vesicles for Measuring Activity of Phospholipase C Isozymes

    • Adam J. Carr, Edhriz Siraliev-Perez, Weigang Huang, John Sondek, Qisheng Zhang
    Pages 225-236

  19. Back Matter

    Pages 237-239
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Keywords

About this book

This detailed book examines experimental approaches used to investigate the regulation and function of phosphoinositides (PtdInsP), rare eukaryotic phospholipids with a broad role in biological processes such as signal transduction, cell migration and adhesion, cell growth, subcellular organization, and membrane trafficking. The combination of complementary biochemical, mass spectrometry, and imaging methods are instrumental for the detection and quantification of PtdInsP species, as well as induced dimerization methods, affinity precipitation or co-sedimentation with liposomes, protein insertion within lipid bilayers, and enzymatic assays, or through emerging methods like native mass spectrometry and microfluidics, all of which are touched upon in this volume. Written in the highly successful Methods in Molecular Biology format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratoryprotocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and practical, Phosphoinositides: Methods and Protocols provides detailed methodology for both specialist and novice researchers on a variety of complementary methods that have been instrumental in dissecting the regulation, dynamics, and function of PtdInsPs.

Editors and Affiliations

  • Department of Chemistry and Biology and the Graduate Program in Molecular Science, Ryerson University, Toronto, Canada

    Roberto J. Botelho

Bibliographic Information

  • Book Title: Phosphoinositides

  • Book Subtitle: Methods and Protocols

  • Editors: Roberto J. Botelho

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-1142-5

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2021

  • Hardcover ISBN: 978-1-0716-1141-8Published: 23 January 2021

  • Softcover ISBN: 978-1-0716-1144-9Published: 23 January 2022

  • eBook ISBN: 978-1-0716-1142-5Published: 22 January 2021

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 239

  • Number of Illustrations: 7 b/w illustrations, 43 illustrations in colour

  • Topics: Lipidology, Cell Biology

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