The use of styrene-maleic acid copolymer (SMA) for studies on T cell membrane rafts

0501169 - ÚMG 2020 RIV NL eng J - Článek v odborném periodiku
Angelisová, Pavla - Ballek, Ondřej - Sýkora, Jan - Benada, Oldřich - Čajka, Tomáš - Pokorná, Jana - Pinkas, Dominik - Hořejší, Václav
The use of styrene-maleic acid copolymer (SMA) for studies on T cell membrane rafts.
Biochimica Et Biophysica Acta-Biomembranes. Roč. 1861, č. 1 (2019), s. 130-141. ISSN 0005-2736. E-ISSN 1879-2642
Grant CEP: GA ČR(CZ) GA17-05903S; GA MŠMT(CZ) LO1509; GA MŠMT(CZ) LM2015062; GA ČR(CZ) GBP302/12/G101; GA MŠMT(CZ) EF16_013/0001775
Institucionální podpora: RVO:68378050 ; RVO:67985823 ; RVO:61388971 ; RVO:61388955
Klíčová slova: Membrane rafts * sma * GPI-anchored proteins * T lymphocytes * Src family kinases * Membrane proteins
Obor OECD: Biochemistry and molecular biology; Physical chemistry (UFCH-W); Microbiology (MBU-M); Analytical chemistry (FGU-C)
Impakt faktor: 3.411, rok: 2019
Způsob publikování: Open access
https://doi.org/10.1016/j.bbamem.2018.08.006

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.
Trvalý link: http://hdl.handle.net/11104/0293384