Journal of Biochemistry Advance Access originally published online on April 19, 2008
Journal of Biochemistry 2008 144(2):167-176; doi:10.1093/jb/mvn056
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© 2008 The Japanese Biochemical Society
Susceptibilities of Phospholipid Membranes Containing Cholesterol or Ergosterol to Gramicidin and its Derivative Incorporated in Lysophospholipid Micelles
1Department of Environmental Sciences, Faculty of Science, Osaka Women's University; and 2Department of Biological Science, Graduate School of Science, Osaka Prefecture University, Japan
*To whom correspondence should be addressed. Tel: and Fax: +81-72-254-9185, E-mail: onda{at}b.s.osakafu-u.ac.jp
Received March 29, 2008; Accepted April 8, 2008
| Abstract |
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Complex formation of gramicidin (GA) and desformylgramicidin (des-GA) with sterols was investigated by measuring the intrinsic Trp fluorescence. In organic solvents, the Trp fluorescence of momeric GA was quenched upon binding either cholesterol or ergosterol, but that of monomeric des-GA was not quenched by adding cholesterol. Both dimeric GA and des-GA bound highly to ergosterol, but not to cholesterol, determined by quenching of Trp fluorescence. Furthermore, GA- and des-GA-loaded lysophosphatidylcholine micelles were incubated with phosphatidylcholine vesicles containing cholesterol or ergosterol. The results showed that both monomeric and dimeric peptides hardly bound to cholesterol incorporated into phospholipid vesicles, but markedly bound to ergosterol incorporated into the bilayer membranes. Interestingly, des-GA bound more specifically to the two sterols than GA. In addition, fluorescence resonance energy transfer analysis showed that des-GA bound more specifically to the two sterol than GA.
Key Words: drug delivery systems, fluorescence, gramicidin, Lysophospholipid, sterol
Abbreviations: CMC, critical micelle concentration; des-GA, desformylgramicidin; DMSO, dimethyl sulphoxide; DPPC, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine; eggPC, egg-yolk phosphatidylcholine; FRET, fluorescence resonance energy transfer; GA, gramicidin; LPC, lysophosphatidylcholine; NATA, N-acetyltryptophanamide; THF, tetrahydrofuran; TNS, 2-(p-toluidino)naphthalene-6-sulphonic acid