Abstract
Apidaecin peptides from Apis mellifera hemolymph are believed to attack intracellular bacterial targets. Our in vivo results for apidaecins 1a and 1b confirm that bacterial activity is non-lytic, however, the manner in which these peptides pass through the cell membrane to exert this activity is unknown. These data are combined with fluorescence (dye leakage) and quartz crystal microbalance studies to investigate the membrane interaction for these two wildtype peptides. It was found that the peptides penetrate the membrane in a trans-membrane manner. The amount of peptide uptake by the membrane is proportional to the concentration of the peptide, however, this appears to be a dynamic equilibrium which can be almost completely reversed by addition of buffer medium. Interestingly, a small residual mass remains within the membrane and the amount of peptide remaining in the membrane is a function of the buffer-salt concentration viz. in high salt, the residual peptide mass remaining is small whereas at low salt concentration, a larger mass of peptide remains bound. These results support a direct membrane penetration mechanism by the wild type apidaecins 1a and 1b. In both cases the peptide–membrane interaction has a negligible effect on the membrane, although, in high salt a permanent change in the membrane does occur at the highest peptide concentration which does not recover following peptide removal.
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Abbreviations
- MPA:
-
Mercaptopropionic acid
- SAM:
-
Self-assembled monolayer
- MIC:
-
Minimal inhibitory concentration
- TSB:
-
Tryptic soy broth
- QCM:
-
Quartz crystal microbalance
- TFA:
-
Trifluoroacetic acid
- ONPG:
-
o-nitrophenyl-β-galactoside
- DMPC:
-
1,2-dimyristoyl-sn-glycero-3-(phosphocholine)
- DMPG:
-
1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] sodium salt
- PBS:
-
Phosphate buffered saline
- DPPC:
-
Dipalmitoylphosphatidylcholine
- SUVs:
-
Small unilamellar vesicles
- Tris-HCl:
-
Tris(hydroxymethyl)aminomethane hydrochloride
- ∆f :
-
Frequency change
- ∆D :
-
Dissipation change
- EDTA:
-
Ethylenediaminetetracetic acid
- RP-HPLC:
-
Reverse phase-high performance liquid chromatography
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Acknowledgments
We acknowledge the financial support from the Australian Research Council Discovery Grant DP0662816 to LLM and AM, Monash Fellowship to AM and the support of Mr Paul Barrett, ATA Scientific. This work was supported by the European Fund for Regional Structure Development (EFRE, European Union and Free State Saxony) to RH.
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Stefania Piantavigna and Patricia Czihal contributed equally to this article.
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Piantavigna, S., Czihal, P., Mechler, A. et al. Cell Penetrating Apidaecin Peptide Interactions with Biomimetic Phospholipid Membranes. Int J Pept Res Ther 15, 139–146 (2009). https://doi.org/10.1007/s10989-009-9175-2
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DOI: https://doi.org/10.1007/s10989-009-9175-2