Membrane docking mode of the C2 domain of PKCε: An infrared spectroscopy and FRET study
| Autor: | Juan C. Gómez-Fernández, Senena Corbalán-García, Alessio Ausili, Antonio L. Egea-Jiménez, Mattias Berglin, Hans Elwing |
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| Rok vydání: | 2013 |
| Předmět: |
Models
Molecular Adenosine Spectrophotometry Infrared Protein Conformation Lipid Bilayers Molecular Conformation ATR-IR Biophysics Synthetic membrane Glycerophospholipids Protein Kinase C-epsilon Biochemistry Protein Structure Secondary chemistry.chemical_compound Fluorescence Resonance Energy Transfer Humans C2 domains Protein–lipid interaction Lipid bilayer POPC Phospholipids C2 domain PKCε Models Statistical Chemistry Phosphatidylethanolamines Cell Biology Lipids Enzyme structure Protein Structure Tertiary Membrane docking Crystallography Membrane Mitochondrial Membranes Phosphatidylcholines Calcium lipids (amino acids peptides and proteins) Protein Binding |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Biomembranes. 1828:552-560 |
| ISSN: | 0005-2736 |
| Popis: | The C2 domain of PKCε binds to negatively charged phospholipids but little is known so far about the docking orientation of this domain when it is bound. By using a FRET assay we have studied the binding of this domain to model membranes. We have also used ATR-Fourier transform infrared spectroscopy with polarized light (ATR-FTIR) to determine the docking mode by calculating the β-sandwich orientation when the domain is bound to different types of model membranes. The vesicle lipid compositions were: POPC/POPE/POPA (22:36:42) imitating the inner leaflet of a plasma membrane, POPC/POPA (50:50) in which POPE has been eliminated with respect to the former composition and POPC/POPE/CL (43:36:21) imitating the inner mitochondrial membrane. Results show that the β-sandwich of the PKCα-C2 domain is inclined at an angle α close to 45° to the membrane normal. Some differences were found with respect to the extent of binding as a function of phospholipid composition and small changes on secondary structure were only evident when the domain was bound to model membranes of POPC/POPA: in this case, the percentage of β-sheet of the C2 domain increases if compared with the secondary structure of the domain in the absence of vesicles. With respect to the β-sandwich orientation, when the domain is bound to POPC/POPE/CL membranes it forms an angle with the normal to the surface of the lipid bilayer (39°) smaller than that one observed when the domain interacts with vesicles of POPC/POPA (49°). |
| Databáze: | OpenAIRE |
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