Overproduction of human Zip (SLC39) zinc transporters in saccharomyces cerevisiae for biophysical characterization
(2021) In Cells 10(2). p.1-21- Abstract
Zinc constitutes the second most abundant transition metal in the human body, and it is implicated in numerous cellular processes, including cell division, DNA and protein synthesis as well as for the catalytic activity of many enzymes. Two major membrane protein families facilitate zinc homeostasis in the animal kingdom, i.e., Zrt/Irt‐like proteins (ZIPs aka solute carrier 39, SLC39, family) and Zn transporters (ZnTs), essentially conducting zinc flux in the opposite directions. Human ZIPs (hZIPs) regulate import of extracellular zinc to the cytosol, being critical in preventing overaccumulation of this potentially toxic metal, and crucial for diverse physiological and pathological processes, including development of neurodegenerative... (More)
Zinc constitutes the second most abundant transition metal in the human body, and it is implicated in numerous cellular processes, including cell division, DNA and protein synthesis as well as for the catalytic activity of many enzymes. Two major membrane protein families facilitate zinc homeostasis in the animal kingdom, i.e., Zrt/Irt‐like proteins (ZIPs aka solute carrier 39, SLC39, family) and Zn transporters (ZnTs), essentially conducting zinc flux in the opposite directions. Human ZIPs (hZIPs) regulate import of extracellular zinc to the cytosol, being critical in preventing overaccumulation of this potentially toxic metal, and crucial for diverse physiological and pathological processes, including development of neurodegenerative disorders and several cancers. To date, our understanding of structure–function relationships governing hZIP‐mediated zinc transport mechanism is scarce, mainly due to the notorious difficulty in overproduction of these proteins for biophysical characterization. Here we describe employment of a Saccharomyces cerevisiae‐based platform for heterologous expression of hZIPs. We demonstrate that yeast is able to produce four full‐length hZIP members belonging to three different subfamilies. One target (hZIP1) is purified in the high quantity and homogeneity required for the downstream biochemical analysis. Our work demonstrates the potential of the described production system for future structural and functional studies of hZIP transporters.
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- author
- Becares, Eva Ramos ; Pedersen, Per Amstrup ; Gourdon, Pontus LU and Gotfryd, Kamil
- organization
- publishing date
- 2021-02-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Family, Membrane proteins, Overproduction, Production platform, Protein purification, Saccharomyces cerevisiae, SLC39, Solute carrier 39, Yeast, Zinc, Zinc transporters, ZIPs
- in
- Cells
- volume
- 10
- issue
- 2
- article number
- 213
- pages
- 21 pages
- publisher
- MDPI AG
- external identifiers
-
- pmid:33494457
- scopus:85100483801
- ISSN
- 2073-4409
- DOI
- 10.3390/cells10020213
- language
- English
- LU publication?
- yes
- id
- 83632ec1-cd42-4708-88a0-80cba154baaa
- date added to LUP
- 2022-03-22 15:32:27
- date last changed
- 2024-06-07 01:55:35
@article{83632ec1-cd42-4708-88a0-80cba154baaa,
abstract = {{<p>Zinc constitutes the second most abundant transition metal in the human body, and it is implicated in numerous cellular processes, including cell division, DNA and protein synthesis as well as for the catalytic activity of many enzymes. Two major membrane protein families facilitate zinc homeostasis in the animal kingdom, i.e., Zrt/Irt‐like proteins (ZIPs aka solute carrier 39, SLC39, family) and Zn transporters (ZnTs), essentially conducting zinc flux in the opposite directions. Human ZIPs (hZIPs) regulate import of extracellular zinc to the cytosol, being critical in preventing overaccumulation of this potentially toxic metal, and crucial for diverse physiological and pathological processes, including development of neurodegenerative disorders and several cancers. To date, our understanding of structure–function relationships governing hZIP‐mediated zinc transport mechanism is scarce, mainly due to the notorious difficulty in overproduction of these proteins for biophysical characterization. Here we describe employment of a Saccharomyces cerevisiae‐based platform for heterologous expression of hZIPs. We demonstrate that yeast is able to produce four full‐length hZIP members belonging to three different subfamilies. One target (hZIP1) is purified in the high quantity and homogeneity required for the downstream biochemical analysis. Our work demonstrates the potential of the described production system for future structural and functional studies of hZIP transporters.</p>}},
author = {{Becares, Eva Ramos and Pedersen, Per Amstrup and Gourdon, Pontus and Gotfryd, Kamil}},
issn = {{2073-4409}},
keywords = {{Family; Membrane proteins; Overproduction; Production platform; Protein purification; Saccharomyces cerevisiae; SLC39; Solute carrier 39; Yeast; Zinc; Zinc transporters; ZIPs}},
language = {{eng}},
month = {{02}},
number = {{2}},
pages = {{1--21}},
publisher = {{MDPI AG}},
series = {{Cells}},
title = {{Overproduction of human Zip (SLC39) zinc transporters in saccharomyces cerevisiae for biophysical characterization}},
url = {{http://dx.doi.org/10.3390/cells10020213}},
doi = {{10.3390/cells10020213}},
volume = {{10}},
year = {{2021}},
}