The assembly factor P17 from bacteriophage PRD1 interacts with positively charged lipid membranes.
The interactions of the assembly factor P17 of bacteriophage PRD1 with liposomes were investigated by static light scattering, fluorescence spectroscopy, and differential scanning calorimetry. Our data show that P17 binds to positively charged large unilamellar vesicles composed of the zwitterionic 1-palmitoyl-2-oleoyl-phosphatidylcholine and sphingosine, whereas only a weak interaction is evident for 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles. P17 does not bind to negatively charged membranes composed of 1-palmitoyl-2-oleoyl-phosphatidylglycerol and 1-palmitoyl-2-oleoyl-phosphatidylcholine. Our differential scanning calorimetry results reveal that P17 slightly perturbs the phase behaviour of neutral phosphatidylcholine and negatively charged multilamellar vesicles. In contrast, the phase transition temperature of positively charged dimyristoylphosphatidylcholine/sphingosine multilamellar vesicles (molar ratio 9 : 1, respectively) is increased by approximately 2.4 degrees C and the half width of the enthalpy peak broadened from 1.9 to 5.6 degrees C in the presence of P17 (protein : lipid molar ratio 1 : 47). Moreover, the enthalpy peak is asymmetrical, suggesting that lipid phase separation is induced by P17. Based on the far-UV CD spectra, the alpha-helicity of P17 increases upon binding to positively charged micelles composed of Triton X-100 and sphingosine. We propose that P17 can interact with positively charged lipid membranes and that this binding induces a structural change on P17 to a more tightly packed and ordered structure.
Holopainen
J M
JM
Helsinki Biophysics and Biomembrane Group Department of Medical Chemistry, Institute of Biomedicine, Helsinki, Finland. jmholopa@rock.helsinki.fi
Säily
M
M
Caldentey
J
J
Kinnunen
P K
PK
D002152:Calorimetry, Differential Scanning; D003090:Coliphages / Q000378:metabolism*; D004735:Energy Transfer; D007700:Kinetics; D016350:Leucine Zippers; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D012542:Scattering, Radiation; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000378:metabolism*
D008081:Liposomes; D014759:Viral Envelope Proteins; C118469:assembly factor P17