Inositol phosphates : methods and protocols / edited by Gregory J. Miller. -- New York : Humana Press, c2020. – (58.17/M592/v.2091)

Contents

Preface

Contributors

1  A Short Historical Perspective of Methods in Inositol Phosphate Research

PART I  ISOLATION AND MEASUREMENT

2  Quantitation of Inositol Phosphates by HPLC-ESI-MS

3  Isolation of Inositol Hexakisphosphate from Soils by Alkaline Extraction and Hypobromite Oxidation

4  Detection of Inositol Phosphates by Split PH Domains

5  Dictyostelium discoideum as a Model to Study Inositol Polyphosphates and Inorganic Polyphosphate

PART II  ASSAYS FOR FUNCTION

6  Characterizing Enzymes of the Diphosphoinositol Polyphosphate Phosphohydrolase (DIPP) Family

7  Metabolic Labeling of inositol Phosphates and Phosphatidylinositols in Yeast and Mammalian Cells

8  Back-Pyrophosphorylation Assay to Detect In Vivo InsP7-Dependent Protein Pyrophosphorylation in Mammalian Cells

9  Analyses of Ligand Binding to IP3 Receptors Using Fluorescence Polarization

10  Investigating the InsP3 Receptor in Living Cells by Caged InsP3

11  Optical Measurement of Synaptic Vesicle Fusion and Its Inhibition by Inositol Pyrophosphate in Primary Cultured Hippocampal Neurons

12  Competitive Fluorescent Ligand Assay for Inositol 1,4,5-Trisphosphate

13  Studying the Effects of Inositol Pyrophosphates in an In Vitro Vesicle-Vesicle Fusion Assay

PART HI  CHEMICAL TOOLS

14  Engineering Protease-Resistant and Highly Active Phytases

15  Direct Cloning Method for Expression of Recombinant Proteins with an Inositol Hexakisphosphate Inducible Self-Cleaving Tag

Index