Intracellular Lipid Transport: Methods and Protocols

ISBN-13: 9781493991358 / Angielski / Twarda / 2019 / 345 str.

Guillaume Drin

Spis treści

1.       Advanced quantitative measure of sterol movements in mammal cells (example of STARD protein).  

2.       Use of new probes to detect sphingolipids

3.       How to measure ceramide transport. 

4.       Update on the strategy to follow the movement of DAG and PA lipids.

5.       TIRF microscopy to measure lipid movement in and out the PM with fluorescent-tagged probe

6.       BRET & FRET approaches to measure intracellular lipid transport.

7.       Measure of dynamics of native lipid species (caged-lipids) in cells.

8.       Cell fractionation and lipidomics to measure lipid movements in organelles.

9.       PS and PE transport at ER-mitochondria contact sites and inside mitochondria

10.   Lipid transport and endosomal-lysosomal compartment.

11.   Measurement of neutral lipid and sterol accumulation in lipid droplets.

12.   Lipid transport in plants (ER-chloroplast contact sites).

13.   Measure of glycolipids transport.

14.   Identification of putative lipid-transporter by sequence analysis.

15.   Identification of lipid transporter and their ligand by purification affinity and denaturating mass spectrometry

16.   Use of click-chemistry to identify novel lipid transporters and their ligands.

17.   In vitro real-time measure of lipid exchange by ORP/Osh protein - Guillaume Drin et al.

18.   Measure of lipid transport by liposome separation approaches

19.   In silico examination of membrane binding and lipid release activity of an LTP

20.    Advanced approaches to measure lipid scramblase/flippase activity

21.   Additionnal : mathematical approaches for modeling lipid transport in cells 

Blurb

This detailed book provides technical approaches to tackle a variety of questions related to intracellular lipid transport in order to improve our understanding at different scales of how lipids are accurately displaced between organelles, across long distances or at membrane contact sites, or within cellular membranes. The volume begins with methodologies to measure the movement of varied lipid species between or in organelle membranes, inside eukaryotic cells, including plant cells, or in bacteria, and continues in vitro or in silico approaches aiming to define, more from a biochemical and structural standpoints, how lipid transfer proteins (LTPs) or flippases/scramblases precisely function. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

 Authoritative and practical, Intracellular Lipid Transport: Methods and Protocols serves as an ideal guide for researchers seeking to shed light on diverse aspects of this critical and often elusive cellular process.