TY - JOUR
T1 - Cryo-electron microscopy of coagulation Factor VIII bound to lipid nanotubes
AU - Parmenter, Christopher D.J.
AU - Cane, Matthew C.
AU - Zhang, Rui
AU - Stoilova-McPhie, Svetla
N1 - Funding Information:
The authors are grateful to Baxter USA for providing the rFVIII, to the British Heart Foundation for funding, Grant: PG/04/070 and The Wellcome trust (UK) Grant 055663/Z/98/Z supporting the EM facility at the Department of Biological Sciences, University of Warwick. Thanks also go to Dr. Geoffrey Kemball-Cook for continuous support and collaboration on the FVIII project and to Elizabeth Wilson-Kubalek for helpful discussions. This work was also supported by grants to Wah Chiu (NIHP41RR02250 and Robert Welch Foundation).
PY - 2008/2/8
Y1 - 2008/2/8
N2 - Factor VIII (FVIII) is a key protein in blood coagulation, deficiency or malfunction of which causes Haemophilia A. The sole cure for this condition is intravenous administration of FVIII, whose membrane-bound structure we have studied by Cryo-electron microscopy and image analysis. Self-assembled lipid nanotubes were optimised to bind FVIII at close to native conditions. The tubes diameter was constant at 30 nm and the lipid bilayer resolved. The FVIII molecules were well defined, forming an 8.5 nm thick outer layer, and appeared to reach the hydrophobic core of the bilayer. The two known FVIII atomic models were superimposed with the averaged 2D protein densities. The insertion of the FVIII within the membrane was evaluated, reaffirming that the membrane-binding C2 or C1-C2 domain(s) fully penetrate the outer leaflet of the lipid layer. The presented results lay the basis for new models of the FVIII overall orientation and membrane-binding mechanism.
AB - Factor VIII (FVIII) is a key protein in blood coagulation, deficiency or malfunction of which causes Haemophilia A. The sole cure for this condition is intravenous administration of FVIII, whose membrane-bound structure we have studied by Cryo-electron microscopy and image analysis. Self-assembled lipid nanotubes were optimised to bind FVIII at close to native conditions. The tubes diameter was constant at 30 nm and the lipid bilayer resolved. The FVIII molecules were well defined, forming an 8.5 nm thick outer layer, and appeared to reach the hydrophobic core of the bilayer. The two known FVIII atomic models were superimposed with the averaged 2D protein densities. The insertion of the FVIII within the membrane was evaluated, reaffirming that the membrane-binding C2 or C1-C2 domain(s) fully penetrate the outer leaflet of the lipid layer. The presented results lay the basis for new models of the FVIII overall orientation and membrane-binding mechanism.
KW - Coagulation Factor VIII
KW - Cryo-electron microscopy
KW - Image analysis
KW - Lipid nanotubes
KW - Membrane-binding
KW - Models fitting
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U2 - 10.1016/j.bbrc.2007.11.072
DO - 10.1016/j.bbrc.2007.11.072
M3 - Article
C2 - 18039465
AN - SCOPUS:37449003239
SN - 0006-291X
VL - 366
SP - 288
EP - 293
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -