Effect of end group blockage on the properties of a class A amphipathic helical peptide.
In a recent classification of biologically active amphipathic alpha-helixes, the lipid-associating domains in exchangeable plasma apolipoproteins have been classified as class A amphipathic helixes (Segrest, J.P., De Loof, H., Dohlman, J.G., Brouillette, C.G., Anantharamaiah, G.M. Proteins 8:103-117, 1990). A model peptide analog with the sequence, Asp Trp Leu Lys Ala Phe Tyr Asp Lys Val Ala Glu Lys Leu Lys Glu Ala Phe (18A), possesses the characteristics of a class A amphipathic helix. The addition of an acetyl group at the alpha-amino terminus and an amide at the alpha-carboxyl terminus, to obtain Ac-18A-NH2, produces large increases in helicity for the peptide both in solution and when associated with lipid (for 18A vs Ac-18A-NH2, from 6 to 38% helix in buffer and from 49 to 92% helix when bound to dimyristoyl phosphatidylcholine in discoidal complexes). Blocking of the end-groups of 18A stabilizes the alpha-helix in the presence of lipid by approximately 1.3 kcal/mol. There is also an increase in the self-association of the blocked peptide in aqueous solution. The free energy of binding to the PC-water interface is increased only by about 3% (from -8.0 kcal/mol for 18A to -8.3 kcal/mol for Ac-18A-NH2). The Ac-18A-NH2 has a much greater potency in raising the bilayer to hexagonal phase transition temperature of dipalmitoleoyl phosphatidylethanolamine than does 18A. In this regard Ac-18A-NH2 more closely resembles the behavior of the apolipoprotein A-I, which is the major protein component of high-density lipoprotein and a potent inhibitor of lipid hexagonal phase formation. The activation of the plasma enzyme lecithin: cholesterol acyltransferase by the Ac-18A-NH2 peptide is greater than the 18A analog and comparable to that observed with the apo A-I. In the case of Ac-18A-NH2, the higher activating potency may be due, at least in part, to the ability of the peptide to micellize egg PC vesicles.
Venkatachalapathi
Y V
YV
Department of Medicine, University of Alabama School of Medicine, Birmingham 35294.
Phillips
M C
MC
Epand
R M
RM
Epand
R F
RF
Tytler
E M
EM
Segrest
J P
JP
Anantharamaiah
G M
GM
D016632:Apolipoprotein A-I / Q000737:chemistry / Q000378:metabolism; D001053:Apolipoproteins / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D004134:Dimyristoylphosphatidylcholine; D004789:Enzyme Activation; D019791:Guanidine; D006146:Guanidines / Q000737:chemistry; D008027:Light; D008051:Lipid Bilayers; D008055:Lipids / Q000737:chemistry; D008566:Membranes / Q000737:chemistry; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000378:metabolism; D010713:Phosphatidylcholines; D011108:Polymers; D011487:Protein Conformation; D012542:Scattering, Radiation; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013499:Surface Properties
D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D006146:Guanidines; D008051:Lipid Bilayers; D008055:Lipids; D010455:Peptides; D010713:Phosphatidylcholines; D011108:Polymers; D012996:Solutions; C080383:peptide 18A; D007862:Phosphatidylcholine-Sterol O-Acyltransferase; D019791:Guanidine; D004134:Dimyristoylphosphatidylcholine