Ion Channels of Various Types Induced in Lipid Membranes by Gramicidin A Derivatives Carrying a Cationic Sequence at Their C-Termini

T. B. Stoilova^1 #, E. A. Dutseva², A. A. Pashkovskaya², S. V. Sychev¹, S. I. Koval’chuk¹, A. A. Sobko², N. S. Egorova¹, E. A. Kotova², Yu. N. Antonenko², A. Yu. Surovoi¹ and V. T. Ivanov¹

^#Phone: (495) 335-58-44; e-mail: stojlova@yahoo.com

¹ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997, Russia;
² Belozersky Institute of Physicochemical Biology, Moscow State University, Vorob’evy Gory, Moscow, 119992, Russia

Received: October 16, 2006;  in final form: January 15, 2007

Abstract.  The channel-forming activity of gramicidin A derivatives carrying positively charged amino acid sequences at their C-termini was studied on planar bilayer lipid membranes and liposomes. We showed previously (FEBS Lett., 2005, vol. 579, pp. 5247–5252) that, at low concentrations, these peptides form classical cation-selective pores typical of gramicidin A, whereas, at high concentrations, they form large nonselective pores. The ability of the peptides to form nonselective pores, which was determined by the efflux of carboxyfluorescein, an organic dye, from liposomes, decreased substantially as the length of the gramicidin fragment in the series of cationic analogues was truncated. CD spectra showed that large pores are formed by peptides having both β^6.3 single-stranded and β^5.6 double-stranded helical conformations of the gramicidin fragment, with the C-terminal cationic sequence being extended. The dimerization of the peptides by the oxidation of the terminal cysteine promoted the formation of nonselective pores. It was shown that nonselective pores are not formed in membranes of erythrocytes, which may indicate a dependence of the channel-forming ability on the membrane type. The results may be of interest for the directed synthesis of peptides with antibacterial activity.

Key words:  gramicidin, cationic analogues; ion channels, nonselective pores; transmembrane amphipathic peptides

Russian Journal of Bioorganic Chemistry 2007, 33(5): 474-481