Inhibition of Escherichia coli ATP synthase by amphibian antimicrobial peptides.
Previously melittin, the alpha-helical basic honey bee venom peptide, was shown to inhibit F(1)-ATPase by binding at the beta-subunit DELSEED motif of F(1)F(o)-ATP synthase. Herein, we present the inhibitory effects of the basic alpha-helical amphibian antimicrobial peptides, ascaphin-8, aurein 2.2, aurein 2.3, carein 1.8, carein 1.9, citropin 1.1, dermaseptin, maculatin 1.1, maganin II, MRP, or XT-7, on purified F(1) and membrane bound F(1)F(0)Escherichia coli ATP synthase. We found that the extent of inhibition by amphibian peptides is variable. Whereas MRP-amide inhibited ATPase essentially completely (approximately 96% inhibition), carein 1.8 did not inhibit at all (0% inhibition). Inhibition by other peptides was partial with a range of approximately 13-70%. MRP-amide was also the most potent inhibitor on molar scale (IC(50) approximately 3.25 microM). Presence of an amide group at the c-terminal of peptides was found to be critical in exerting potent inhibition of ATP synthase ( approximately 20-40% additional inhibition). Inhibition was fully reversible and found to be identical in both F(1)F(0) membrane preparations as well as in isolated purified F(1). Interestingly, growth of E. coli was abrogated in the presence of ascaphin-8, aurein 2.2, aurein 2.3, citropin 1.1, dermaseptin, magainin II-amide, MRP, MRP-amide, melittin, or melittin-amide but was unaffected in the presence of carein 1.8, carein 1.9, maculatin 1.1, magainin II, or XT-7. Hence inhibition of F(1)-ATPase and E. coli cell growth by amphibian antimicrobial peptides suggests that their antimicrobial/anticancer properties are in part linked to their actions on ATP synthase.
Laughlin
Thomas F
TF
Department of Biological Sciences, Box 70703, East Tennessee State University, Johnson City, TN 37614, USA.
Ahmad
Zulfiqar
Z
D000595:Amino Acid Sequence; D000663:Amphibians / Q000378:metabolism*; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000494:pharmacology*; D025243:Bacterial Proton-Translocating ATPases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000302:isolation & purification; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D003470:Culture Media; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D005947:Glucose / Q000494:pharmacology; D008555:Melitten / Q000737:chemistry / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000187:drug effects
D023181:Antimicrobial Cationic Peptides; D003470:Culture Media; D008555:Melitten; D025243:Bacterial Proton-Translocating ATPases; D005947:Glucose