Inositol phosphates : methods and protocols [electronic resource]

  • Miller, Gregory J.
  • Humana Press
  • 2020
Inositol phosphates : methods and protocols [electronic resource]


This volume describes modern approaches for the detection and analysis of inositol phosphates from various biological, clinical, and environmental samples. Chapters focus on methods to study enzymes that modify inositol signals, chemical biology of inositol phosphates, and methods to synthesize and purify inositol phosphates and their analogs. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls.


Part I: Isolation and Measurement
1. A Short Historical Perspective of Methods in Inositol Phosphate Research
Stephen B. Shears
2. Quantitation of Inositol Phosphates by HPLC-ESI-MS
Quynh H. Duong and Ronald B. Pegg
3. Isolation of Inositol Hexakisphosphate from Soils by Alkaline Extraction and Hypobromite Oxidation
Benjamin L. Turner
4. Detection of Inositol Phosphates by Split PH Domains
Reiko Sakaguchi, Shunsuke Tajima, Yasuo Mori, and Takashi Morii
5. Dictyostelium discoideum as a Model to Study Inositol Pyrophosphate and Inorganic Polyphosphate
Yann Desfougères and Adolfo Saiardi

Part II: Assays for Function
6. Characterising Enzymes of the Diphosphoinositol Polyphosphate Phosphohydrolase (DIPP) Family
Lucinda Winward, Rajagopal Sharada Kilari, and Stephen T. Safrany
7. Metabolic Labeling of Inositol Phosphates and Phosphatidylinositols in Yeast and Mammalian Cells
Andrew T. Hale, Bradley P. Clarke, and John D. York
8. Back-pyrophosphorylation Assay to Detect in vivo Ins7-dependent Protein Pyrophosphorylation in Mammalian Cells
Manasa Chanduri and Rashna Bhandari
9. Analyses of Ligand Binding to IP3 receptors using Fluorescence Polarization
Ana M. Rossi and Colin W. Taylor
10. Investigating the InsP3 Receptor in Living Cells by Caged InsP3
Xin Hui and Peter Lipp
11. Optical Measurement of Synaptic Vesicle Fusion and its Inhibition by Inositol Pyrophosphate in Primary Cultured Hippocampal Neurons
Sung Hyun Kim
12. Competitive Fluorescent Ligand Assay for Inositol 1,4,5-Trisphosphate
Akihiko Tanimura and Satoshi Shuto
13. Studying the Effects of Inositol Pyrophosphates in an In Vitro Vesicle-vesicle Fusion Assay
Mi Sook Sung and Tae-Young Yoon

Part III: Chemical Tools
14. Engineering Protease Resistant and Highly Active Phytases
Canfang Niu, Peilong Yang, and Bin Yao
15. Direct Cloning Method for Expression of Recombinant Proteins with an Inositol Hexakisphosphate Inducible Self-cleaving Tag
Keehwan Kwon, Marco Biancucci, Patrick J. Woida, and Karla J. F. Satchell