Abstract
A number of previous studies have shown that phospholipid molecules come and go between the endoplasmic reticulum (ER) and mitochondrial membranes while the molecular basis of non-vesicular phospholipid transport is still not understood well. In this chapter, we describe an optimized method that uses membrane fractions isolated from yeast cells to directly analyze phospholipid transport between the ER and mitochondria. With this assay, we are able to assess not only the ER-to-mitochondria but also mitochondria-to-ER transports at the same time. We believe that this assay system can accelerate the research on inter-organelle phospholipid trafficking.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Flis VV, Daum G (2013) Lipid transport between the endoplasmic reticulum and mitochondria. Cold Spring Harb Perspect Biol 5:a013235
Tamura Y, Sesaki H, Endo T (2014) Phospholipid transport via mitochondria. Traffic 15:933–945
Tatsuta T, Scharwey M, Langer T (2014) Mitochondrial lipid trafficking. Trends Cell Biol 24:44–52
Shen H, Heacock PN, Clancey CJ et al (1996) The CDS1 gene encoding CDP-diacylglycerol synthase in Saccharomyces cerevisiae is essential for cell growth. J Biol Chem 271:789–795
Clancey CJ, Chang SC, Dowhan W (1993) Cloning of a gene (PSD1) encoding phosphatidylserine decarboxylase from Saccharomyces cerevisiae by complementation of an Escherichia coli mutant. J Biol Chem 268:24580–24590
Nikawa J, Yamashita S (1984) Molecular cloning of the gene encoding CDPdiacylglycerol-inositol 3-phosphatidyl transferase in Saccharomyces cerevisiae. Eur J Biochem 143:251–256
Kodaki T, Yamashita S (1987) Yeast phosphatidylethanolamine methylation pathway. Cloning and characterization of two distinct methyltransferase genes. J Biol Chem 262:15428–15435
Tamura Y, Harada Y, Nishikawa S et al (2013) Tam41 is a CDP-diacylglycerol synthase required for cardiolipin biosynthesis in mitochondria. Cell Metab 17:709–718
Kojima R, Endo T, Tamura Y (2016) A phospholipid transfer function of ER-mitochondria encounter structure revealed in vitro. Sci Rep 6:30777
Kornmann B, Currie E, Collins SR et al (2009) An ER-mitochondria tethering complex revealed by a synthetic biology screen. Science 325:477–481
Kawano S, Tamura Y, Kojima R et al (2018) Structure-function insights into direct lipid transfer between membranes by Mmm1-Mdm12 of ERMES. J Cell Biol 217:959–974
Friedman JR, Kannan M, Toulmay A et al (2018) Lipid homeostasis is maintained by dual targeting of the mitochondrial PE biosynthesis enzyme to the ER. Dev Cell 44:261–270
Acknowledgments
This work was supported by JSPS KAKENHI (Grant Numbers 15H05595 and 17H06414 to Y.T. and 15H05705 and 22227003 to T.E.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Tamura, Y., Kojima, R., Endo, T. (2019). Advanced In Vitro Assay System to Measure Phosphatidylserine and Phosphatidylethanolamine Transport at ER/Mitochondria Interface. In: Drin, G. (eds) Intracellular Lipid Transport. Methods in Molecular Biology, vol 1949. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9136-5_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9136-5_6
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9135-8
Online ISBN: 978-1-4939-9136-5
eBook Packages: Springer Protocols