Helix-helix association of a lipid-bound amphipathic alpha-helix derived from apolipoprotein C-II.
The interaction of a peptide derived from the sequence of apolipoprotein C-II (apoC-II) with a model lipid surface has been investigated by fluorescence spectroscopy. ApoC-II19-39, labeled at the N-terminus with 7-nitrobenz-2-oxa-1,3-diazole (NBD), bound to small unilamellar vesicles of phosphatidylcholine with a dissociation constant of 6 microM. The lipid-bound NBD-labeled peptide exhibited a red-edge excitation shift in its emission maximum and anisotropy, consistent with insertion of the probe into the motionally restricted, polar environment provided by the bilayer interface. The small Stokes shift of the NBD fluorophore permits electronic energy homotransfer between peptides on the lipid surface and results in depolarization of the NBD emission. At high surface densities of lipid-bound peptide, the anisotropy of the NBD probe was 33% lower than in corresponding samples in which electronic energy homotransfer was prevented by the addition of an unlabeled peptide. The efficiency of energy transfer between probes was not consistent with a random distribution of peptides on the lipid surface, indicating instead the self-association of lipid-bound apoC-II19-39. We propose that the role of this sequence in apoC-II is not only to mediate binding of protein to a lipid surface, but also to stabilize the lipoprotein complexes by associating with other amphipathic helices within apoC-II and with other apolipoproteins.
MacPhee
C E
CE
Russell Grimwade School of Biochemistry and Molecular Biology, University of Melbourne, Parkville, Victoria, Australia.
Howlett
G J
GJ
Sawyer
W H
WH
Clayton
A H
AH
D009327:4-Chloro-7-nitrobenzofurazan / Q000737:chemistry; D000595:Amino Acid Sequence; D053304:Apolipoprotein C-II; D001056:Apolipoproteins C / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D004530:Egg Yolk; D004735:Energy Transfer; D005454:Fluorescence Polarization; D007202:Indicators and Reagents; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence
D053304:Apolipoprotein C-II; D001056:Apolipoproteins C; D007202:Indicators and Reagents; D008051:Lipid Bilayers; D010446:Peptide Fragments; D010713:Phosphatidylcholines; D009327:4-Chloro-7-nitrobenzofurazan