Advanced In Vitro Assay System to Measure Phosphatidylserine and Phosphatidylethanolamine Transport at ER/Mitochondria Interface
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.
Key wordsMitochondria Endoplasmic reticulum Phospholipid transport In vitro reconstitution Yeast
This work was supported by JSPS KAKENHI (Grant Numbers 15H05595 and 17H06414 to Y.T. and 15H05705 and 22227003 to T.E.).