A novel heavy-atom label for side-specific peptide iodination: synthesis, membrane incorporation and X-ray reflectivity
| Autor: | André Beerlink, Brigitte Worbs, Tim Salditt, Ulf Diederichsen, Philipp E. Schneggenburger |
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| Rok vydání: | 2009 |
| Předmět: |
Conformational change
Circular dichroism Halogenation Lipid Bilayers Peptide 010402 general chemistry 01 natural sciences Protein Structure Secondary Cell membrane 03 medical and health sciences chemistry.chemical_compound X-Ray Diffraction Metals Heavy medicine Peptide synthesis Physical and Theoretical Chemistry Lipid bilayer 030304 developmental biology chemistry.chemical_classification 0303 health sciences Chemistry Circular Dichroism Cell Membrane Atomic and Molecular Physics and Optics 0104 chemical sciences Amino acid Crystallography medicine.anatomical_structure Membrane Peptides Iodine |
| Zdroj: | Chemphyschem : a European journal of chemical physics and physical chemistry. 10(9-10) |
| ISSN: | 1439-7641 |
| Popis: | Structural parameters, such as conformation, orientation and penetration depth of membrane-bound peptides and proteins that may function as channels, pores or biocatalysts, are of persistent interest and have to be probed in the native fluid state of a membrane. X-ray scattering in combination with heavy-atom labeling is a powerful and highly appropriate method to reveal the position of a certain amino acid residue within a lipid bilayer with respect to the membrane normal axis up to a resolution of several Angstrom. Herein, we report the synthesis of a new iodine-labeled amino acid building block. This building block is intended for peptide incorporation to provide high intensities for electron density difference analysis of X-ray reflectivity data and improve the labeling potential for the lipid bilayer head-group and water region. The novel building block as well as the commercially available non-iodinated analogue, required for X-ray scattering, was implemented in a transmembrane peptide motif via manual solid-phase peptide synthesis (SPPS) following the fluorenylmethyloxycarbonyl (Fmoc)-strategy. The derived peptides were reconstituted in lipid vesicles as well as in highly aligned multilamellar lipid stacks and investigated via circular dichroism (CD) and X-ray reflectivity. Thereby, it has been revealed that the bulky iodine probe neither causes conformational change of the peptide structure nor lamellar disordering of the membrane complexes. |
| Databáze: | OpenAIRE |
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