Abstract
The dynamin-related proteins (DRPs) are self-assembling membrane remodeling machines that are indispensable for fundamental cellular trafficking and homeostatic processes. We describe in this chapter protocols developed in our laboratory for purification of full-length and minimal constructs of Chaetomium thermophilum Vps1, the model fungal DRP, using mammalian and Escherichia coli expression systems.
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References
Ford MGJ, Chappie JS (2019) The structural biology of the dynamin-related proteins: new insights into a diverse, multitalented family. Traffic 20(10):717–740
Praefcke GJ, McMahon HT (2004) The dynamin superfamily: universal membrane tubulation and fission molecules? Nat Rev Mol Cell Biol 5(2):133–147
Hoppins S, Lackner L, Nunnari J (2007) The machines that divide and fuse mitochondria. Annu Rev Biochem 76:751–780
Antonny B, Burd C, De Camilli P et al (2016) Membrane fission by dynamin: what we know and what we need to know. EMBO J 35(21):2270–2284
Faelber K, Gao S, Held M et al (2013) Oligomerization of dynamin superfamily proteins in health and disease. Prog Mol Biol Transl Sci 117:411–443
Varlakhanova NV, Alvarez FJD, Brady TM et al (2018) Structures of the fungal dynamin-related protein Vps1 reveal a unique, open helical architecture. J Cell Biol 217(10):3608–3624
Arlt H, Reggiori F, Ungermann C (2015) Retromer and the dynamin Vps1 cooperate in the retrieval of transmembrane proteins from vacuoles. J Cell Sci 128(4):645–655
Chi RJ, Liu J, West M, Wang J, Odorizzi G, Burd CG (2014) Fission of SNX-BAR-coated endosomal retrograde transport carriers is promoted by the dynamin-related protein Vps1. J Cell Biol 204(5):793–806
Hayden J, Williams M, Granich A et al (2013) Vps1 in the late endosome-to-vacuole traffic. J Biosci 38(1):73–83
Kuravi K, Nagotu S, Krikken AM et al (2006) Dynamin-related proteins Vps1p and Dnm1p control peroxisome abundance in Saccharomyces cerevisiae. J Cell Sci 119(Pt 19):3994–4001
Peters C, Baars TL, Buhler S, Mayer A (2004) Mutual control of membrane fission and fusion proteins. Cell 119(5):667–678
Smaczynska-de R II, Allwood EG, Aghamohammadzadeh S, Hettema EH, Goldberg MW, Ayscough KR (2010) A role for the dynamin-like protein Vps1 during endocytosis in yeast. J Cell Sci 123(Pt 20):3496–3506
Acknowledgement
We thank Tyler M. Brady and Bryan A. Tornabene for assistance with protein purification. This work was supported by the National Institutes of Health grant GM120102 (M. G. J. Ford).
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Varlakhanova, N.V., Ford, M.G.J. (2020). Purification of the Dynamin-Related Protein Vps1 Using Mammalian and Bacterial Expression Systems. In: Ramachandran, R. (eds) Dynamin Superfamily GTPases. Methods in Molecular Biology, vol 2159. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0676-6_2
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DOI: https://doi.org/10.1007/978-1-0716-0676-6_2
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