The use of styrene-maleic acid copolymer (SMA) for studies on T cell membrane rafts
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
30463696
DOI
10.1016/j.bbamem.2018.08.006
PII: S0005-2736(18)30237-2
Knihovny.cz E-resources
- Keywords
- GPI-anchored proteins, Membrane proteins, Membrane rafts, SMA, Src family kinases, T lymphocytes,
- MeSH
- Anisotropy MeSH
- Cell Membrane chemistry MeSH
- Cholesterol chemistry MeSH
- Detergents chemistry MeSH
- Chromatography, Gel MeSH
- Jurkat Cells MeSH
- Humans MeSH
- Lipid Bilayers chemistry MeSH
- Lipids chemistry MeSH
- Maleates chemistry MeSH
- Fatty Acids chemistry MeSH
- Membrane Microdomains chemistry MeSH
- Membrane Proteins chemistry MeSH
- Membranes, Artificial MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Polymers chemistry MeSH
- Scattering, Radiation MeSH
- Solubility MeSH
- Styrene chemistry MeSH
- Light MeSH
- T-Lymphocytes cytology MeSH
- Ultracentrifugation MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Cholesterol MeSH
- Detergents MeSH
- Lipid Bilayers MeSH
- Lipids MeSH
- Maleates MeSH
- maleic acid MeSH Browser
- Fatty Acids MeSH
- Membrane Proteins MeSH
- Membranes, Artificial MeSH
- Polymers MeSH
- Styrene MeSH
An emerging alternative to the use of detergents in biochemical studies on membrane proteins is apparently the use styrene-maleic acid (SMA) amphipathic copolymers. These cut the membrane into nanodiscs (SMA-lipid particles, SMALPs), which contain membrane proteins possibly surrounded by their native lipid environment. We examined this approach for studies on several types of T cell membrane proteins, previously defined as raft or non-raft associated, to see whether the properties of the raft derived SMALPs differ from non-raft SMALPs. Our results indicate that two types of raft proteins, GPI-anchored proteins and two Src family kinases, are markedly present in membrane fragments much larger (>250 nm) than those containing non-raft proteins (<20 nm). Lipid probes sensitive to membrane fluidity (membrane order) indicate that the lipid environment in the large SMALPs is less fluid (more ordered) than in the small ones which may indicate the presence of a more ordered lipid Lo phase which is characteristic of membrane rafts. Also the lipid composition of the small vs. large SMALPs is markedly different - the large ones are enriched in cholesterol and lipids containing saturated fatty acids. In addition, we confirm that T cell membrane proteins present in SMALPs can be readily immunoisolated. Our results support the use of SMA as a potentially better (less artifact prone) alternative to detergents for studies on membrane proteins and their complexes, including membrane rafts.
Institute of Physiology of the Czech Academy of Sciences Vídeňská 1083 142 20 Praha 4 Czech Republic
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