Competitive interactions of amphipathic polycationic peptides and cationic fluorescent probes with lipid membrane: experimental approaches and computational model.
The electrostatic interaction of polycationic peptides with negatively charged biomembranes has been recognized as the first and very important step of their selective binding to many bacteria and transformed cells. In this work we demonstrated the phenomenon of competition of some earlier designed polycationic peptides and fluorescent probes for their binding to the negatively charged inner membrane of mitochondria and to the PC/PG (9:1) liposomes. Rat liver mitochondria swelling induced by the antimicrobial polycationic peptide BTM-P1 (VAPIAKYLATALAKWALKQGFAKLKS) and by the retro-BTM-P1 was significantly diminished in the presence of 10μM fluorescent probe safranin O. In experiments with liposomes, the polycationic peptides BTM-P1 and P7-5 (IYLATALAKWALKQGF-GG-RRRRRRR) at the concentrations of 2-3μM completely displaced the membrane-bound fluorescent probe DiSC3(5) in a low ionic strength medium. The developed computational model allowed a mathematical description of such interactions, predicting membrane surface concentrations of bound peptides as the function of the membrane surface charge and lipid quantity in the sample, the peptide charge, hydrophobicity and concentration, the ionic strength of incubation medium and of the presence of a charged fluorescent probe used for monitoring the membrane surface potential under real-time peptide-membrane interactions.
Escuela de Física, Facultad de Ciencias, Universidad Nacional de Colombia, Sede Medellín, Calle 59A, No 63-20, Medellín, Colombia. Electronic address: vvasilie@unal.edu.co.
Victor V
VV
Lemeshko
D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002412:Cations / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D051381:Rats
D023181:Antimicrobial Cationic Peptides; C501422:BTM-P1 peptide; D002412:Cations; D005456:Fluorescent Dyes; D008081:Liposomes
Computational model; Fluorescent probes; Liposomes; Mitochondria; Peptide–membrane interaction; Polycationic peptides