title: Effects of increasing hydrophobicity on the physical-chemical and biological properties of a class A amphipathic helical peptide.
abstract: We have recently shown that a class A amphipathic peptide 5F with increased amphipathicity protected mice from diet-induced atherosclerosis (Garber et al. J. Lipid Res. 2001. 42: 545-552). We have now examined the effects of increasing the hydrophobicity of a series of homologous class A amphipathic peptides, including 5F, on physical and functional properties related to atherosclerosis inhibition by systematically replacing existing nonpolar amino acids with phenylalanine. The peptides, based on the sequence Ac-D-W-L-K-A-F-Y-D-K-V-A-E-K-L-K-E-A-F-NH(2) (Ac-18A-NH(2) or 2F) were: 3F(3)(Ac-F(3)18A-NH(2)), 3F(14)(Ac-F(14)18A-NH(2)), 4F(Ac-F(3,14)18A-NH(2)), 5F(Ac-F(11,14,17) 18A-NH(2)), 6F(Ac-F(10,11,14,17)18A-NH(2)), and 7F(Ac-F(3,10,11,14,17) 18A-NH(2)). Measurements of aqueous solubility, HPLC retention time, exclusion pressure for penetration into an egg phosphatidylcholine (EPC) monolayer, and rates of EPC solubilization revealed an abrupt increase in the hydrophobicity between peptides 4F and 5F; this was accompanied by increased ability to associate with phospholipids. The peptides 6F and 7F were less effective, indicating a limit to increased hydrophobicity for promoting lipid interaction in these peptides. Despite this marked increase in lipid affinity, these peptides were less effective than apoA-I in activating the plasma enzyme, lecithin:cholesterol acyltransferase, with 5F activating LCAT the best (80% of apoA-I). Peptides 4F, 5F, and 6F were equally potent in inhibiting LDL-induced monocyte chemotactic activity. These studies suggest that an appropriate balance between peptide-peptide and peptide-lipid interactions is required for optimal biological activity of amphipathic peptides. These studies provide a rationale for the design of small apoA-I-mimetics with increased potency for atherosclerosis inhibition.
authors.affiliation: The Atherosclerosis Research Unit and the Departments of Medicine and Biochemistry and Molecular Genetics, The University of Alabama at Birmingham Medical Center, Birmingham, AL 35294, USA.
authors.forename: G
authors.initials: G
authors.lastname: Datta
authors.forename: M
authors.initials: M
authors.lastname: Chaddha
authors.forename: S
authors.initials: S
authors.lastname: Hama
authors.forename: M
authors.initials: M
authors.lastname: Navab
authors.forename: A M
authors.initials: AM
authors.lastname: Fogelman
authors.forename: D W
authors.initials: DW
authors.lastname: Garber
authors.forename: V K
authors.initials: VK
authors.lastname: Mishra
authors.forename: R M
authors.initials: RM
authors.lastname: Epand
authors.forename: R F
authors.initials: RF
authors.lastname: Epand
authors.forename: S
authors.initials: S
authors.lastname: Lund-Katz
authors.forename: M C
authors.initials: MC
authors.lastname: Phillips
authors.forename: J P
authors.initials: JP
authors.lastname: Segrest
authors.forename: G M
authors.initials: GM
authors.lastname: Anantharamaiah
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001665:Binding Sites / Q000502:physiology; D002478:Cells, Cultured; D002633:Chemotaxis / Q000187:drug effects; D002784:Cholesterol / Q000378:metabolism; D008078:Cholesterol, LDL / Q000494:pharmacology; D002942:Circular Dichroism; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D051379:Mice; D009000:Monocytes / Q000502:physiology; D010455:Peptides / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000187:drug effects* / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism; D012995:Solubility
chemical_list: D016632:Apolipoprotein A-I; D008078:Cholesterol, LDL; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D010713:Phosphatidylcholines; D010743:Phospholipids; C582295:apolipoprotein A-I mimetic peptide 4F; C431648:peptide 5F; D002784:Cholesterol; D007862:Phosphatidylcholine-Sterol O-Acyltransferase