| Autor: |
Bürck J; Institute of Biological Interfaces (IBG-2), Karlsruhe Institute of Technology (KIT) , POB 3640, D-76021 Karlsruhe, Germany., Wadhwani P; Institute of Biological Interfaces (IBG-2), Karlsruhe Institute of Technology (KIT) , POB 3640, D-76021 Karlsruhe, Germany., Fanghänel S; Institute of Biological Interfaces (IBG-2), Karlsruhe Institute of Technology (KIT) , POB 3640, D-76021 Karlsruhe, Germany., Ulrich AS; Institute of Biological Interfaces (IBG-2), Karlsruhe Institute of Technology (KIT) , POB 3640, D-76021 Karlsruhe, Germany.; Institute of Organic Chemistry, KIT , Fritz-Haber-Weg 6, D-76131 Karlsruhe, Germany. |
| Jazyk: |
angličtina |
| Zdroj: |
Accounts of chemical research [Acc Chem Res] 2016 Feb 16; Vol. 49 (2), pp. 184-92. Date of Electronic Publication: 2016 Jan 12. |
| DOI: |
10.1021/acs.accounts.5b00346 |
| Abstrakt: |
The structures of membrane-bound polypeptides are intimately related to their functions and may change dramatically with the lipid environment. Circular dichroism (CD) is a rapid analytical method that requires relatively low amounts of material and no labeling. Conventional CD is routinely used to monitor the secondary structure of peptides and proteins in solution, for example, in the presence of ligands and other binding partners. In the case of membrane-active peptides and transmembrane proteins, these measurements can be applied to, and remain limited to, samples containing detergent micelles or small sonicated lipid vesicles. Such traditional CD analysis reveals only secondary structures. With the help of an oriented circular dichroism (OCD) setup, however, based on the preparation of macroscopically oriented lipid bilayers, it is possible to address the membrane alignment of a peptide in addition to its conformation. This approach has been mostly used for α-helical peptides so far, but other structural elements are conceivable as well. OCD analysis relies on Moffitt's theory, which predicts that the electronic transition dipole moments of the backbone amide bonds in helical polypeptides are polarized either parallel or perpendicular to the helix axis. The interaction of the electric field vector of the circularly polarized light with these transitions results in an OCD spectrum of a membrane-bound α-helical peptide, which exhibits a characteristic line shape and reflects the angle between the helix axis and the bilayer normal. For parallel alignment of a peptide helix with respect to the membrane surface (S-state), the corresponding "fingerprint" CD band around 208 nm will exhibit maximum negative amplitude. If the helix changes its alignment via an obliquely tilted (T-state) to a fully inserted transmembrane orientation (I-state), the ellipticity at 208 nm decreases and the value approaches zero due to the decreased interactions between the field and the transition dipole. Compared to conventional CD, OCD data are not only collected in the biologically relevant environment of a highly hydrated planar lipid bilayer (whose composition can be varied at will), but in addition it provides information about the tilt angle of the polypeptide in the membrane. It is the method of choice for screening numerous different conditions, such as peptide concentration, lipid composition, membrane additives, pH, temperature, and sample hydration. All these factors have been found to affect the peptide alignment in membrane, while having little or no influence on conformation. In many cases, the observed realignment could be related to biological action, such as pore formation by antimicrobial and cell-penetrating peptides, or to binding events of transmembrane segments of integral membrane proteins. Likewise, any lipid-induced conversion from α-helix to β-sheeted conformation is readily picked up by OCD and has been interpreted in terms of protein instability or amyloid-formation. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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