The use of pore-forming toxins to image lipids and lipid domains.
Fiche publication
Date publication
janvier 2021
Journal
Methods in enzymology
Auteurs
Membres identifiés du Cancéropôle Est :
Pr MELY Yves, Dr RICHERT Ludovic
Tous les auteurs :
Tomishige N, Murate M, Didier P, Richert L, Mély Y, Kobayashi T
Lien Pubmed
Résumé
Very few proteins are reported to bind specific lipids. Because of the high selectivity and strong binding to specific lipids, lipid-targeting pore forming toxins (PFTs) have been employed to study the distribution of lipids in cell- and model-membranes. Non-toxic and monomeric PFT-derivatives are especially useful to study living cells. In this chapter we highlight sphingomyelin (SM)-binding PFT, lysenin (Lys), its derivatives, and newly identified SM/cholesterol binding protein, nakanori. We describe the preparation of non-toxic mutant of Lys (NT-Lys) and its application in optical and super resolution microscopy. We also discuss the observation of nanometer scale lipid domains labeled with nakanori and maltose-binding protein (MBP)-Lys in electron microscopy.
Mots clés
Cholesterol, Electron microscopy, Freeze fracture replica, Lipid asymmetry, Lipid-binding protein, Lysenin, Sphingomyelin, Super resolution microscopy
Référence
Methods Enzymol. 2021 ;649:503-542