Phosphoinositides: Methods and Protocols

ISBN-13: 9781071611449 / Angielski / Miękka / 2022

Botelho, Roberto J.

Opis

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 laboratory protocols, 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.

Spis treści

1. Simultaneous Detection of Phosphoinositide Lipids by Radioactive Metabolic Labeling

            Noah Steinfeld, Sai Srinivas Panapakkam Giridharan, Emily J. Kauffman, and Lois S. Weisman

 

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

            Avishek Ghosh and Padinjat Raghu

 

3. 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, and Bernard Payrastre

 

4. Quantification of Genetically-Encoded Lipid Biosensors

            Rachel C. Wills, Jonathan Pacheco, and Gerald R.V. Hammond

 

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

            Rebecca Cabral-Dias, Yasmin Awadeh, Roberto J. Botelho, and Costin N. Antonescu

 

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

            Fan Fan, Chen Ji, and Xuelin Lou

 

7. Induced Dimerization Tools to Deplete Specific Phosphatidylinositol Phosphates

            Jonathan Pacheco, Rachel C. Wills, and Gerald R.V. Hammond

 

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

            Taki Nishimura

 

9. Assessing In Situ Phosphoinositide-Protein Interactions through Fluorescence Proximity Ligation Assay in Cultured Cells

            Mo Chen, Hudson T. Horn, Tianmu Wen, Vincent L. Cryns, and Richard A. Anderson

 

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

            Calvin Yeager, Djoshkun Shengjuler, Simou Sun, Paul S. Cremer, and Craig E. Cameron

 

11. Exploring Phosphoinositide Binding Using Native Mass Spectrometry

            Julian Bender and Carla Schmidt

 

12. Liposome-Based Methods to Study Protein-Phosphoinositide Interaction

            Mélanie Mansat, Mélanie Picot, Gaëtan Chicanne, Virginie Nahoum, Frédérique Gaits-Iacovoni, Bernard Payrastre, Karim Hnia, and Julien Viaud

 

13. Liposome-Based Methods to Study GTPases Activation by Phosphoinositides

            Julien Viaud, Laurie Ceccato, Bernard Payrastre, and Frédérique Gaits-Iacovoni

 

14. Liposome Co-Sedimentation and Co-Flotation Assays to Study Lipid-Protein Interactions

            Yosuke Senju, Pekka Lappalainen, and Hongxia Zhao

 

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

            Ángel Pérez-Lara and Reinhard Jahn

 

16. Fluorescence Assays to Study Membrane Penetration of Proteins

            Yosuke Senju and Hongxia Zhao

 

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

            Adam J. Carr, Edhriz Siraliev-Perez, Weigang Huang, John Sondek, and Qisheng Zhang