Microfluidic-Derived Detection of Protein-Facilitated Copper Flux Across Lipid Membranes
(2022) In Analytical Chemistry 94(34). p.11831-11837- Abstract
Measurement of protein-facilitated copper flux across biological membranes is a considerable challenge. Here, we demonstrate a straightforward microfluidic-derived approach for visualization and measurement of membranous Cu flux. Giant unilamellar vesicles, reconstituted with the membrane protein of interest, are prepared, surface-immobilized, and assessed using a novel quencher-sensor reporter system for detection of copper. With the aid of a syringe pump, the external buffer is exchanged, enabling consistent and precise exchange of solutes, without causing vesicle rupture or uneven local metal concentrations brought about by rapid mixing. This approach bypasses common issues encountered when studying heavy metal-ion flux, thereby... (More)
Measurement of protein-facilitated copper flux across biological membranes is a considerable challenge. Here, we demonstrate a straightforward microfluidic-derived approach for visualization and measurement of membranous Cu flux. Giant unilamellar vesicles, reconstituted with the membrane protein of interest, are prepared, surface-immobilized, and assessed using a novel quencher-sensor reporter system for detection of copper. With the aid of a syringe pump, the external buffer is exchanged, enabling consistent and precise exchange of solutes, without causing vesicle rupture or uneven local metal concentrations brought about by rapid mixing. This approach bypasses common issues encountered when studying heavy metal-ion flux, thereby providing a new platform for in vitro studies of metal homeostasis aspects that are critical for all cells, health, and disease.
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- author
- Górecki, Kamil LU ; Hansen, Jesper S. LU ; Li, Ping LU ; Nayeri, Niloofar LU ; Lindkvist-Petersson, Karin LU and Gourdon, Pontus LU
- organization
- publishing date
- 2022-08-30
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical Chemistry
- volume
- 94
- issue
- 34
- pages
- 7 pages
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- scopus:85136535455
- pmid:35969432
- ISSN
- 0003-2700
- DOI
- 10.1021/acs.analchem.2c02081
- language
- English
- LU publication?
- yes
- id
- e67ed40d-e88f-4de3-b7ca-e0dad805f1be
- date added to LUP
- 2022-10-17 09:53:42
- date last changed
- 2024-04-18 04:55:11
@article{e67ed40d-e88f-4de3-b7ca-e0dad805f1be,
abstract = {{<p>Measurement of protein-facilitated copper flux across biological membranes is a considerable challenge. Here, we demonstrate a straightforward microfluidic-derived approach for visualization and measurement of membranous Cu flux. Giant unilamellar vesicles, reconstituted with the membrane protein of interest, are prepared, surface-immobilized, and assessed using a novel quencher-sensor reporter system for detection of copper. With the aid of a syringe pump, the external buffer is exchanged, enabling consistent and precise exchange of solutes, without causing vesicle rupture or uneven local metal concentrations brought about by rapid mixing. This approach bypasses common issues encountered when studying heavy metal-ion flux, thereby providing a new platform for in vitro studies of metal homeostasis aspects that are critical for all cells, health, and disease.</p>}},
author = {{Górecki, Kamil and Hansen, Jesper S. and Li, Ping and Nayeri, Niloofar and Lindkvist-Petersson, Karin and Gourdon, Pontus}},
issn = {{0003-2700}},
language = {{eng}},
month = {{08}},
number = {{34}},
pages = {{11831--11837}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Analytical Chemistry}},
title = {{Microfluidic-Derived Detection of Protein-Facilitated Copper Flux Across Lipid Membranes}},
url = {{http://dx.doi.org/10.1021/acs.analchem.2c02081}},
doi = {{10.1021/acs.analchem.2c02081}},
volume = {{94}},
year = {{2022}},
}