CPL - Chalmers Publication Library
| Utbildning | Forskning | Styrkeområden | Om Chalmers | In English In English Ej inloggad.

Assigning Membrane Binding Geometry of Cytochrome c by Polarized Light Spectroscopy

Christina Caesar (Institutionen för kemi- och bioteknik, Fysikalisk kemi ; SuMo Biomaterials) ; Elin Esbjörner (Institutionen för kemi- och bioteknik, Fysikalisk kemi) ; Per Lincoln (Institutionen för kemi- och bioteknik, Fysikalisk kemi) ; Bengt Nordén (Institutionen för kemi- och bioteknik, Fysikalisk kemi)
Biophysical Journal (0006-3495). Vol. 96 (2009), 8, p. 3399-3411.
[Artikel, refereegranskad vetenskaplig]

In this work we demonstrate how polarized light absorption spectroscopy (linear dichroism (LD)) analysis of the peptide ultraviolet-visible spectrum of a membrane-associated protein (cytochrome (cyt) c) allows orientation and structure to be assessed with quite high accuracy in a native membrane environment that can be systematically varied with respect to lipid composition. Cyt c binds strongly to negatively charged lipid bilayers with a distinct orientation in which its a-helical segments are on average parallel to the membrane surface. Further information is provided by the LID of the pi-pi* transitions of the heme porphyrin and transitions of aromatic residues, mainly a single tryptophan. A good correlation with NMR data was found, and combining NMR structural data with LID angular data allowed the whole protein to be docked to the lipid membrane. When the redox state of cyt c was changed, distinct variations in the LID spectrum of the heme Soret band were seen corresponding to changes in electronic transition energies; however, no significant change in the overall protein orientation or structure was observed. Cyt c is known to interact in a specific manner with the doubly negatively charged lipid cardiolipin, and incorporation of this lipid into the membrane at physiologically relevant levels was indeed found to affect the protein orientation and its a-helical content. The detail in which cyt c binding is described in this study shows the potential of LID spectroscopy using shear-deformed lipid vesicles as a new methodology for exploring membrane protein structure and orientation.

Nyckelord: protein secondary structure, circular-dichroism spectra, optical, rotatory dispersion, linear dichroism, ferricytochrome-c, mitochondrial-membrane, peripheral proteins, simple polypeptides, lipid-membranes, flow dichroism

Denna post skapades 2010-01-14. Senast ändrad 2016-03-31.
CPL Pubid: 107246


Läs direkt!

Länk till annan sajt (kan kräva inloggning)

Institutioner (Chalmers)

Institutionen för kemi- och bioteknik, Fysikalisk kemi (2005-2014)
SuMo Biomaterials


Fysikalisk kemi

Chalmers infrastruktur