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The use of styrene-maleic acid copolymer (SMA) for studies on T cell membrane rafts
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SYSNO ASEP 0501169 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title The use of styrene-maleic acid copolymer (SMA) for studies on T cell membrane rafts Author(s) Angelisová, Pavla (UMG-J) RID
Ballek, Ondřej (UMG-J)
Sýkora, Jan (UFCH-W) RID
Benada, Oldřich (MBU-M) ORCID, RID
Čajka, Tomáš (FGU-C) RID, ORCID, SAI
Pokorná, Jana (UMG-J)
Pinkas, Dominik (UMG-J) ORCID
Hořejší, Václav (UMG-J) RIDNumber of authors 8 Source Title Biochimica Et Biophysica Acta-Biomembranes. - : Elsevier - ISSN 0005-2736
Roč. 1861, č. 1 (2019), s. 130-141Number of pages 12 s. Publication form Print - P Language eng - English Country NL - Netherlands Keywords Membrane rafts ; sma ; GPI-anchored proteins ; T lymphocytes ; Src family kinases ; Membrane proteins Subject RIV EB - Genetics ; Molecular Biology OECD category Biochemistry and molecular biology Subject RIV - cooperation J. Heyrovsky Institute of Physical Chemistry - Physical ; Theoretical Chemistry
Institute of Microbiology - Microbiology, Virology
Institute of Physiology - Analytical Chemistry, SeparationR&D Projects GA17-05903S GA ČR - Czech Science Foundation (CSF) LO1509 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LM2015062 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GBP302/12/G101 GA ČR - Czech Science Foundation (CSF) EF16_013/0001775 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UMG-J - RVO:68378050 ; FGU-C - RVO:67985823 ; MBU-M - RVO:61388971 ; UFCH-W - RVO:61388955 UT WOS 000451494500015 EID SCOPUS 85051646885 DOI 10.1016/j.bbamem.2018.08.006 Annotation 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. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2020 Electronic address https://doi.org/10.1016/j.bbamem.2018.08.006
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