期刊
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
卷 1861, 期 1, 页码 130-141出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbamem.2018.08.006
关键词
Membrane rafts; SMA; GPI-anchored proteins; T lymphocytes; Src family kinases; Membrane proteins
资金
- Czech Science Foundation [17-05903S, P302-12-G101]
- Institute of Molecular Genetics of the Czech Academy of Sciences [RVO 68378050]
- Ministry of Education, Youth and Sports (MEYS) of the Czech Republic [LO1509]
- Institute of Physiology of the Czech Academy of Sciences [RVO:67985823]
- Czech-Biolmaging large RI project - MEYS CR [LM2015062]
- OP RDE [CZ.02.1.01/0.0/0.0/16013/0001775]
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 L-o 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.
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