[Antimicrobial peptides as efficient DNA vectors].
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Anti-Bacterial Agents↗
Biomedical subjects
Burkhard Bechinger
Publications and source records attributed to Burkhard Bechinger.
Magainin 2 in phospholipid bilayers: peptide orientation and lipid chain ordering studied by X-ray diffraction.
We present a structural study of biomimetic lipid bilayers interacting with the antimicrobial peptide magainin 2 amide, using grazing incidence X-ray diffraction and reciprocal space mapping (RSM) techniques. The short-range order of lipid chains in lecithin is found to be strongly reduced by the peptides. From the scattering intensity of the chain correlation peak, we can quantify the lateral length scale R over which the bilayer structure is affected by peptide binding. The non-local perturbation of the bilayer is discussed in the framework of bilayer elasticity theory.
Amino Acid Sequence↗
Guanidine hydrochloride induced equilibrium unfolding studies of colicin B and its channel-forming fragment.
The conformational stabilities of full-length colicin B and its isolated C-terminal domain were studied by guanidine hydrochloride induced unfolding. The unfolding/refolding was monitored by far-UV CD and intrinsic tryptophan fluorescence spectroscopies. At pH 7.4, the disruption of the secondary structure of full-length colicin B is monophasic, while changes in tertiary structure occur in two separate transitions. The intermediate species, which is well-populated around 2.2 M guanidine hydrochloride, exhibits secondary and tertiary structures distinct from both native and unfolded states. Whereas the domain structure of native full-length colicin B is reflected in its DSC profile, the folding intermediate of the same protein exhibits a single unresolved peak. These observations have led us to propose an unfolding model for full-length colicin B where the first transition between 0 and 2.5 M GuHCl with an associated free energy of 3 kcal/mol correlates with the partial unfolding of the R/T domain. The stability of full-length colicin B is weakened due to the presence of the R/T domain in both the native [Ortega, A., Lambotte, S., and Bechinger, B. (2001) J. Biol. Chem. 276 (17), 13563-13572] and the intermediate states. The second transition between 2.5 and 5 M GuHCl involves unfolding of the C-terminal domain (Delta = 7 kcal/mol). The isolated colicin B C-terminal domain consists of two subdomains, and the two parts of this protein fragment unfold sequentially through the formation of at least one intermediate. The significance of these results for membrane insertion of colicin B is discussed.
Calorimetry, Differential Scanning↗
Bilayer sample for fast or slow magic angle oriented sample spinning solid-state NMR spectroscopy.
An alternative setup for Magic Angle Oriented Spinning Spectroscopy is proposed. Samples were prepared by orienting lipid bilayers onto polymer films, which were wrapped into a spiral so as to fit into 4 or 7 mm MAS rotors. This geometry resulted in narrow line widths and a higher upper spinning limit when compared to the conventional MAOSS setup with stacked glass plates. Whereas orientational information was extracted from low spinning spectra, fast spinning will be applicable to high-resolution multidimensional NMR pulse sequences.
Lipid Bilayers↗
A spectroscopic study of the membrane interaction of the antimicrobial peptide Pleurocidin.
The cationic amphipathic alpha-helical antibiotic peptide, pleurocidin, from the winter flounder Pleuronectes americanus associates strongly with anionic membranes where it is able to translocate across the membrane, cause dye leakage from vesicles and induce pore like channel conductance. To investigate the mechanism of pleurocidin antibiotic activity in more detail we have applied a variety of spectroscopic techniques to study the interaction of pleurocidin with model membranes. At neutral pH the peptide inserts into membranes containing anionic lipids and, as shown by proton-decoupled 15N solid-state NMR spectroscopy of macroscopically oriented samples, is aligned parallel to the membrane surface. 2H solid-state NMR spectroscopy of chain deuterated phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) lipids in mixed membranes shows that pleurocidin interacts with both the zwitterionic PE and anionic PG but disrupts the lipid acyl chain order of the anionic PG lipids more effectively. At acidic pH the three histidine residues of pleurocidin become protonated and positively charged which does not alter the membrane disrupting effect nor the location of the peptide in the membrane. The results are interpreted in terms of a structural model for pleurocidin inserted into anionic lipid membranes and the implications of our data are discussed in terms of a general mechanism for the antibiotic activity.
Amino Acid Sequence↗
Translocation of amino acyl residues from the membrane interface to the hydrophobic core: thermodynamic model and experimental analysis using ATR-FTIR spectroscopy.
The interactions of a series of histidine-containing peptides with biological model membranes have been investigated by attenuated total reflection Fourier transform infra red (ATR-FTIR) spectroscopy. Related peptides have previously been shown to exhibit antibiotic and DNA transfection activities. The 26-residue LAH4X4 peptides were designed in such a manner to form amphipathic helical structures in membrane environments. Four histidines and four variable amino acids X constitute one face of the helix whereas leucines and alanines characterize the opposite hydrophobic surface. The dichroic ratio of ATR-FTIR spectra has been used to follow the pH-dependent transition from in-plane to transmembrane alignments upon increase in pH. A theoretical model of the topological modulations is presented and the experimental transition curves analysed in order to reveal the Gibbs free energy of transition. The novel concept provides access to the free energy changes associated with the amino acids X incorporated into an extended alpha-helix and in the context of phospholipid bilayers. For the peptides of the series the Gibbs free energies associated with the transition from the membrane interface to the bilayer interior follow the sequence of amino acids: L<A approximately I<S approximately F<T approximately G<V approximately W<<Y.
Amino Acids↗