Protocol for determining the regulation of lipid kinases and changes in phospholipids in vitro

Cansu Karabiyik; David C. Rubinsztein

STAR Protocols (Dec 2021)

Protocol for determining the regulation of lipid kinases and changes in phospholipids in vitro

Cansu Karabiyik,
David C. Rubinsztein

Affiliations

Cansu Karabiyik: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK; UK Dementia Research Institute, Cambridge, UK; Mortimer B. Zuckerman Mind Brain and Behavior Institute, Columbia University, 3227 Broadway, Manhattan, NY 10027, USA; Corresponding author
David C. Rubinsztein: Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge CB2 0XY, UK; UK Dementia Research Institute, Cambridge, UK; Corresponding author

Journal volume & issue: Vol. 2, no. 4
p. 100926

Abstract

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Summary: The regulation of lipid kinases has remained elusive given the difficulties of assessing changes in lipid levels. Here, we describe the isolation of protein and lipid kinases to determine the regulation of lipid kinases in vitro. This can be followed by analysis of effects of regulators on lipid kinase-mediated changes in phospholipids without the use of radioactivity, with a specific focus on PI(5)P generation by the enzyme PIKfyve.For complete details on the use and execution of this protocol, please refer to Karabiyik et al. (2021).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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