Secondary conformation of short lysine- and leucine-rich peptides assessed by optical spectroscopies: effect of chain length, concentration, solvent, and time.
Solution secondary structures of three synthetic cationic peptides, currently used in antisense oligonucleotide delivery into living cells, have been analyzed by means of circular dichroism (CD) and Raman scattering in different buffers as a function of concentration and time. All three peptides are of minimalist conception, i.e., formed by only two types of amino acids (leucine: L and lysine: K). Two of these peptides contain 15 aminoacids: N(ter)- KLLKLLLKLLLKLLK (L(10)K(5)), N(ter)-KLKLKLKLKLKLKLK (L(7)K(8)), and the third one has only 9 residues: N(ter)-KLKLKLKLK (L(4)K(5)). The conformational behavior of the 15-mers in pure water differs considerably one from another. Although both of them are initially disordered in the 50-350 microM range, L(10)K(5) gradually undergoes a disordered to alpha-helix transition for molecular concentrations above 100 microM. In all other solvents used, L(10)K(5) adopts a stable alpha-helical conformation. In methanol and methanol/Tris mixture, nonnative alpha-helices can be induced in both KL-alternating peptides, i.e., L(7)K(8) and L(4)K(5). However, in major cases and with a time delay depending on peptide concentration, beta-like structures can be gradually formed in both solutions. In PBS and methanol/PBS mixture, the tendency for L(7)K(8) and L(4)K(5) is to form structures belonging to beta-family. A discussion has been undertaken on the effect of counterions as well as their nature in the stabilization of ordered structures in both KL-alternating peptides.
Hernández
Belén
B
UMR CNRS 7033, BioMoCeTi, Université Pierre et Marie Curie, Paris, France.
Boukhalfa-Heniche
F-Z
FZ
Seksek
Olivier
O
Coïc
Yves-Marie
YM
Ghomi
Mahmoud
M
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D007700:Kinetics; D007930:Leucine / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry*; D000432:Methanol / Q000737:chemistry; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000737:chemistry*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012997:Solvents / Q000737:chemistry*; D013057:Spectrum Analysis; D013059:Spectrum Analysis, Raman; D014325:Tromethamine / Q000737:chemistry
D009842:Oligopeptides; D010455:Peptides; D012997:Solvents; D014325:Tromethamine; D007930:Leucine; D008239:Lysine; D000432:Methanol