Isolation and thermal stabilization of mouse ferroportin.

Autor: Deshpande CN; Structural Biology Program, Centenary Institute, Sydney, NSW, Australia., Azucenas CR; Department of Pharmacology & Systems Physiology, University of Cincinnati College of Medicine, OH.; Medical Sciences Baccalaureate Program, University of Cincinnati College of Medicine, OH, USA., Qiao B; Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA, USA., Nomura N; Department of Cell Biology, Graduate School of Medicine, Kyoto University, Japan., Xin V; Structural Biology Program, Centenary Institute, Sydney, NSW, Australia., Font J; Structural Biology Program, Centenary Institute, Sydney, NSW, Australia., Iwata S; Department of Cell Biology, Graduate School of Medicine, Kyoto University, Japan., Ganz T; Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA, USA.; Department of Pathology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA., Nemeth E; Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA, USA., Mackenzie B; Medical Sciences Baccalaureate Program, University of Cincinnati College of Medicine, OH, USA.; Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA, USA., Jormakka M; Structural Biology Program, Centenary Institute, Sydney, NSW, Australia.
Jazyk: angličtina
Zdroj: FEBS open bio [FEBS Open Bio] 2021 Jan; Vol. 11 (1), pp. 26-34. Date of Electronic Publication: 2020 Dec 17.
DOI: 10.1002/2211-5463.13039
Abstrakt: Ferroportin (Fpn) is an essential mammalian iron transporter that is negatively regulated by the hormone hepcidin. Our current molecular understanding of Fpn-mediated iron efflux and regulation is limited due to a lack of biochemical, biophysical and high-resolution structural studies. A critical step towards understanding the transport mechanism of Fpn is to obtain sufficient quantities of pure and stable protein for downstream studies. As such, we detail here an expression and purification protocol for mouse Fpn yielding milligram quantities of pure protein. We have generated deletion constructs exhibiting enhanced thermal stability and which retained iron-transport activity and hepcidin responsiveness, providing a platform for further biophysical studies of Fpn.
(© 2020 The Authors. FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)
Databáze: MEDLINE
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