ROUCHIDANE EYITAYO, Akande; BOUDIER-LEMOSQUET, Axel; CHAIGNEPAIN, Stephane; PRIAULT, Muriel; MANON, Stephen

doi:10.3390/biom13060876

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ROUCHIDANE EYITAYO, Akande
Institut de biochimie et génétique cellulaires [IBGC]

BOUDIER-LEMOSQUET, Axel
Institut de biochimie et génétique cellulaires [IBGC]

CHAIGNEPAIN, Stephane
Chimie et Biologie des Membranes et des Nanoobjets [CBMN]

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Biomolecules. 2023, vol. 13, n° 6, p. 876

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The antiapoptotic protein Bcl-xL is a major regulator of cell death and survival, but many aspects of its functions remain elusive. It is mostly localized in the mitochondrial outer membrane (MOM) owing to its C-terminal hydrophobic α-helix. In order to gain further information about its membrane organization, we set up a model system combining cell-free protein synthesis and nanodisc insertion. We found that, contrary to its proapoptotic partner Bax, neosynthesized Bcl-xL was spontaneously inserted into nanodiscs. The deletion of the C-terminal α-helix of Bcl-xL prevented nanodisc insertion. We also found that nanodisc insertion protected Bcl-xL against the proteolysis of the 13 C-terminal residues that occurs during expression of Bcl-xL as a soluble protein in E. coli. Interestingly, we observed that Bcl-xL increased the insertion of Bax into nanodiscs, in a similar way to that which occurs in mitochondria. Cell-free synthesis in the presence of nanodiscs is, thus, a suitable model system to study the molecular aspects of the interaction between Bcl-xL and Bax during their membrane insertion.< Leer menos

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apoptosis

Bcl-2 family

nanodiscs

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Bcl-xL Is Spontaneously Inserted into Preassembled Nanodiscs and Stimulates Bax Insertion in a Cell-Free Protein Synthesis System

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