Organelle-Selective Labeling of Choline-Containing Phospholipids (CPLs) and Real-Time Imaging in Living Cells

Masaki Tsuchiya; Tomonori Tamura; Itaru Hamachi

doi:10.1002/cpz1.105

Organelle-Selective Labeling of Choline-Containing Phospholipids (CPLs) and Real-Time Imaging in Living Cells

Curr Protoc. 2021 Apr;1(4):e105. doi: 10.1002/cpz1.105.

Authors

Masaki Tsuchiya^{1

2}, Tomonori Tamura¹, Itaru Hamachi^{1

3}

Affiliations

¹ Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto, Japan.
² PRESTO (Precursory Research for Embryonic Science and Technology, JST), 4-1-8 Honcho, Kawaguchi-shi, Saitama, Japan.
³ ERATO (Exploratory Research for Advanced Technology, JST), Sanbancho, Chiyodaku, Tokyo, Japan.

PMID: 33886163
DOI: 10.1002/cpz1.105

Abstract

Choline-containing phospholipids (CPLs), including phosphatidylcholine (PC) and sphingomyelin (SM), are the major components of mammalian cell membranes and play critical roles during a variety of cellular processes. However, intracellular dynamics of CPLs is poorly understood due to a lack of methods to trace CPL trafficking at organelle resolution. Here, we describe protocols that make it possible to fluorescently label CPLs at the targeted organelles and to monitor their movement within living cells using confocal microscopy. © 2021 Wiley Periodicals LLC. Basic Protocol 1: ER-Golgi-selective labeling of azide-tagged CPLs for confocal imaging Basic Protocol 2: Mitochondria-selective labeling of azide-tagged CPLs for confocal imaging.

Keywords: choline-containing phospholipids; click chemistry; live-cell imaging; organelle-selective labeling.

MeSH terms

Animals
Choline*
Golgi Apparatus
Organelles
Phosphatidylcholines
Phospholipids*

Substances

Phosphatidylcholines
Phospholipids
Choline