Detection of Isolated Mitochondria-Associated ER Membranes Using the Sigma-1 Receptor
The interface between the endoplasmic reticulum (ER) and mitochondria referred to as the MAM (mitochondria-associated ER membrane) plays important roles in many physiological functions. A specific marker for this important entity of cellular structure is urgently needed. Thus, we propose in this method chapter that the membrane-bound ER chaperone sigma-1 receptor serves as an ideal marker for the MAM. We describe in detail the preparation and purification of the MAM by using the sigma-1 receptor as the marker and demonstrate the uniqueness of this marker by using a variety of cells, peripheral and neuronal.
Key wordsSigma-1 receptor Chaperone Endoplasmic reticulum Mitochondria MAM SDS/PAGE electrophoresis Fractionation
This work is supported by the Intramural Research Program of the National Institute on Drug Abuse, National Institutes of Health, of the Department of Health and Human Services of the United States of America.
- 9.Ja L, Tata JR (1973) A rapidly sedimenting fraction of rat liver endoplasmic reticulum. J Cell Sci 13:447–459Google Scholar
- 14.Sebastián D, Hernández-Alvarez MI, Segalés J, Sorianello E, Muñoz JP, Sala D, Waget A, Liesa M, Paz JC, Gopalacharyulu P, Matej O, Pich S, Burcelin R, Palacín M, Zorzano A (2012) Mitofusin 2 (Mfn2) links mitochondrial and endoplasmic reticulum function with insulin signaling and is essential for normal glucose homeostasis. Proc Natl Acad Sci U S A 109:5523–5528PubMedPubMedCentralCrossRefGoogle Scholar
- 17.Tubbs E, Theurey P, Vial G, Bendridi N, Bravard A, Chauvin M, Ji-Cao J, Zoulim F, Bartosch B, Ovize M, Vidal H, Rieusset J (2014) Mitochondria-associated endoplasmic reticulum membrane (MAM) integrity is required for insulin signaling and is implicated in hepatic insulin resistance. Diabetes 63:3279–3294PubMedCrossRefGoogle Scholar