title: Lecithin:cholesterol acyltransferase activation by synthetic amphipathic peptides.
abstract: The amphipathic helical theory of Segrest and colleagues (FEBS Lett.:38:247-253, 1974) proposes that the lipid-binding segments of serum apolipoproteins are in an alpha helical conformation. Furthermore the helices have a hydrophobic face and a hydrophilic face with a specific distribution of positively and negatively charged residues. The importance of the pattern of the charged residues in the lipid binding and lecithin:cholesterol acyltransferase (LCAT) activation by the segments is still debated. We designed a 30-residue peptide, GALA, which in the alpha helical conformation has a hydrophilic face composed of glutamic acid residues (Sabbarao et al.: Biochemistry 26:2964-2972, 1987). GALA behaves like the serum apolipoproteins in its interaction with dimyristoylphosphatidylcholine (DMPC) at neutral pH; the amino terminal tryptophan of GALA undergoes a blue shift in its fluorescence emission spectrum, and the circular dichroism (CD) spectrum indicates that GALA acquires alpha helical structure in the presence of DMPC. A DMPC-GALA:19/1 (molar ratio) complex can be isolated by gel-permeation chromatography. This complex has a discoidal structure with the approximate dimensions of 44-A edge thickness and a 170- to 350-A diameter. GALA activates LCAT with DMPC but not with unsaturated phospholipids as the substrate. The apparent partition coefficient of GALA into DMPC vesicles is 100-fold larger than into egg phosphatidylcholine vesicles. The interaction of GALA with unsaturated lipids at neutral pH is so weak that no detectable change in the spectroscopic properties of GALA or the structure of the liposomes can be detected under the conditions used here. The sequence of GALA differs from previously studied model Apo A1 peptides by the absence of positively charged residues on the hydrophilic face. This indicates that positive charges in Apo A1-like peptides are not required in order to form discoidal structures with saturated phospholipids or to activate LCAT with such lipid substrates.
authors.affiliation: Department of Pharmacy, School of Pharmacy, University of California, San Francisco 94143.
authors.forename: N K
authors.initials: NK
authors.lastname: Subbarao
authors.forename: C J
authors.initials: CJ
authors.lastname: Fielding
authors.forename: R L
authors.initials: RL
authors.lastname: Hamilton
authors.forename: F C
authors.initials: FC
authors.lastname: Szoka
mesh_terms: D002850:Chromatography, Gel; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine / Q000302:isolation & purification; D004789:Enzyme Activation / Q000187:drug effects; D008027:Light; D008081:Liposomes; D008854:Microscopy, Electron; D010455:Peptides / Q000494:pharmacology*; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013501:Surface-Active Agents / Q000494:pharmacology*; D014364:Tryptophan
chemical_list: D008081:Liposomes; D010455:Peptides; D010743:Phospholipids; D013501:Surface-Active Agents; D014364:Tryptophan; D007862:Phosphatidylcholine-Sterol O-Acyltransferase; D004134:Dimyristoylphosphatidylcholine