Isolation and thermal stabilization of mouse ferroportin

Isolation and thermal stabilization of mouse ferroportin.

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Item Type: Article
Status: Published
Official URL: https://doi.org/10.1002/2211-5463.13039
Journal or Publication Title: FEBS Open Bio
Volume: 11
Number: 1
Page Range: pp. 26-34
Date: 2021
Divisions: Structural Biology
Depositing User: General Admin
Identification Number: 10.1002/2211-5463.13039
ISSN: 2211-5463
Date Deposited: 07 Jan 2021 01:06
Abstract:

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.

Keywords: ferroportin; hepcidin regulation; iron metabolism; thermostabilization.

© 2020 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.

Creators:
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Deshpande, Chandrika N.
UNSPECIFIED
Azucenas, Corbin R.
UNSPECIFIED
Qiao, Bo
UNSPECIFIED
Nomura, Norimichi
UNSPECIFIED
Xin, Vicky
UNSPECIFIED
Font, Josep
UNSPECIFIED
Iwata, So
UNSPECIFIED
Ganz, Tomas
UNSPECIFIED
Nemeth, Elizabeta
UNSPECIFIED
Mackenzie, Bryan
UNSPECIFIED
Jormakka, Mika
UNSPECIFIED
Last Modified: 07 Jan 2021 01:06
URI: https://eprints.centenary.org.au/id/eprint/936

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