Photolabelling of D-beta-hydroxybutyrate apodehydrogenase with azidoaryl phospholipids.
Two synthetic photoactive azidoarylphosphatidylcholines were used to investigate the level of interaction between D-beta-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.
el Kebbaj
M S
MS
Berrez
J M
JM
Lakhlifi
T
T
Morpain
C
C
Latruffe
N
N
D000345:Affinity Labels / Q000378:metabolism*; D000818:Animals; D001051:Apoenzymes / Q000378:metabolism*; D001059:Apoproteins / Q000378:metabolism*; D001386:Azides / Q000378:metabolism*; D003574:Cytochrome c Group / Q000378:metabolism; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism; D004789:Enzyme Activation; D006884:Hydroxybutyrate Dehydrogenase / Q000378:metabolism*; D008244:Lysophosphatidylcholines / Q000378:metabolism; D008930:Mitochondria, Liver / Q000201:enzymology; D010713:Phosphatidylcholines / Q000378:metabolism*; D010777:Photochemistry; D051381:Rats
D000345:Affinity Labels; D001051:Apoenzymes; D001059:Apoproteins; D001386:Azides; D003574:Cytochrome c Group; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; C035255:myristoyllysophosphatidylcholine; C044850:1-myristoyl-2-(12-N-(4-azido-2-nitrophenyl)aminododecanoyl)phosphatidylcholine; C044851:1-myristoyl-2-(2-azido-4-nitrobenzoyl)phosphatidylcholine; D006884:Hydroxybutyrate Dehydrogenase; D004134:Dimyristoylphosphatidylcholine