Methods to visualise zinc transporter proteins of the SLC39A family in cells [Chapter 4]

Farr, Georgia, Jones, Samuel and Taylor, Kathryn M. ORCID: https://orcid.org/0000-0002-9576-9490 2023. Methods to visualise zinc transporter proteins of the SLC39A family in cells [Chapter 4]. Hu, Jian, ed. Methods in Enzymology, Vol. 687. Elsevier, pp. 67-85. (10.1016/bs.mie.2023.04.020)

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Abstract

Zinc is essential to many important biological processes and the SLC39A family of zinc transporters which help to control zinc levels in cells, are increasingly being implicated in disease states. In order to determine their exact roles in these processes, reliable methods for their successful production are now required. Unfortunately, extensive post-translational modification and temporally specific activation makes visualisation of ZIP family transporters difficult. As such, modifications of common molecular cell biology techniques are necessary to maximise success when studying these proteins. These include zinc stimulation and nocodazole synchronisation to enhance the activation of ZIP7 and ZIP6/ZIP10, respectively. Maximal ZIP6/ZIP10 retention can also be achieved through careful handling of loosely adhered mitotic fractions when harvesting cell cultures for lysis. Transfection can also be used to enhance ZIP visualisation, however consideration of transfection periods and inclusion of sodium butyrate are recommended to enhance transfection efficiency. When probing for ZIP family transporters, we recommend that epitope choice considers post-translational cleavage and phosphorylated protein isoforms. Finally, where expression of a particular ZIP transporter is manipulated, researchers should consider parallel evaluation of related ZIP transporter expression, to account for transporter compensation.

Item Type:	Book Section
Date Type:	Publication
Status:	Published
Schools:	Pharmacy
Publisher:	Elsevier
ISBN:	9780323957762
ISSN:	0076-6879
Date of First Compliant Deposit:	20 October 2023
Date of Acceptance:	9 May 2023
Last Modified:	26 Oct 2023 10:30
URI:	https://orca.cardiff.ac.uk/id/eprint/163336

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