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Photolabelling of D‐β‐hydroxybutyrate apodehydrogenase with azidoaryl phospholipids
Author(s) -
El Kebbaj M'Hamed S.,
Berrez Jean-Marc,
Lakhlifi Tahar,
Morpain Claude,
Latruffe Norbert
Publication year - 1985
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(85)81178-9
Subject(s) - chemistry , phosphatidylcholine , phospholipid , myristoylation , phosphocholine , amphiphile , moiety , lipid bilayer , cofactor , lecithin , stereochemistry , biochemistry , membrane , biophysics , organic chemistry , enzyme , biology , copolymer , polymer
Two synthetic photoactive azidoarylphosphatidylcholines were used to investigate the level of interaction between D‐β‐hydroxybutyrate apodehydrogenase (apoBDH), an amphipathic membrane protein, with the hydrophobic domain of phospholipids. The two synthetic lecithins, PL I (1‐myristoyl‐2‐12‐ N ‐(4‐azido‐2‐nitrophenyl) aminododecanoyl‐ sn ‐glycero‐3‐phosphocholine) and PL II (1‐myristoyl‐2‐(2‐azido‐4‐nitrobenzoyl)‐ sn ‐glycero‐3‐phosphocholine), are able to reactivate the non‐active purified apoBDH as well as the non‐photoactive homologs, indicating that the photoreactive chemical groups are without effect on the cofactor properties of phosphatidylcholine. Photoirradiation of reconstituted complexes between phospholipid containing azidoaryllecithin and apoBDH leads to a covalent binding of some synthetic lecithin molecules on the protein. The labelling, about 3 times higher with PL II than with PL I, suggests that the area of interacting domain of BDH with the hydrophobic moiety of phospholipid is more important at or near the surface of the lipid bilayer than in the inner part. This approach is further demonstration that BDH is an integral protein.