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Biochim Biophys Acta Biomembr. 2017 Nov;1859(11):2155-2160. doi: 10.1016/j.bbamem.2017.08.010. Epub 2017 Aug 25.

Solubilization of human cells by the styrene-maleic acid copolymer: Insights from fluorescence microscopy.

Author information

1
Membrane Biochemistry and Biophysics, Bijvoet Center for Biomolecular Research, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. Electronic address: j.m.dorr@uu.nl.
2
Cell Biology, Department of Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
3
Membrane Biochemistry and Biophysics, Bijvoet Center for Biomolecular Research, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. Electronic address: j.a.killian@uu.nl.

Abstract

Extracting membrane proteins from biological membranes by styrene-maleic acid copolymers (SMAs) in the form of nanodiscs has developed into a powerful tool in membrane research. However, the mode of action of membrane (protein) solubilization in a cellular context is still poorly understood and potential specificity for cellular compartments has not been investigated. Here, we use fluorescence microscopy to visualize the process of SMA solubilization of human cells, exemplified by the immortalized human HeLa cell line. Using fluorescent protein fusion constructs that mark distinct subcellular compartments, we found that SMA solubilizes membranes in a concentration-dependent multi-stage process. While all major intracellular compartments were affected without a strong preference, plasma membrane solubilization was found to be generally slower than the solubilization of organelle membranes. Interestingly, some plasma membrane-localized proteins were more resistant against solubilization than others, which might be explained by their presence in specific membrane domains with differing properties. Our results support the general applicability of SMA for the isolation of membrane proteins from different types of (sub)cellular membranes.

KEYWORDS:

Cellular localization; Membrane domain; Native nanodiscs; Plasma membrane organization; Preferential solubilization; SMA-resistant membranes

PMID:
28847501
DOI:
10.1016/j.bbamem.2017.08.010
[Indexed for MEDLINE]

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