Abstract
Membrane-bound Factor VIII (FVIII) has a critical function in blood coagulation as the pro-cofactor to the serine-protease Factor IXa (FIXa) in the FVIIIa-FIXa complex assembled on the activated platelet membrane. Defects or deficiency of FVIII cause Hemophilia A, a mild to severe bleeding disorder. Despite existing crystal structures for FVIII, its membrane-bound organization has not been resolved. Here we present the dimeric FVIII membrane-bound structure when bound to lipid nanotubes, as determined by cryo-electron microscopy. By combining the structural information obtained from helical reconstruction and single particle subtomogram averaging at intermediate resolution (15-20 Å), we show unambiguously that FVIII forms dimers on lipid nanotubes. We also demonstrate that the organization of the FVIII membrane-bound domains is consistently different from the crystal structure in solution. The presented results are a critical step towards understanding the mechanism of the FVIIIa-FIXa complex assembly on the activated platelet surface in the propagation phase of blood coagulation.
| Original language | English (US) |
|---|---|
| Article number | 11212 |
| Journal | Scientific Reports |
| Volume | 5 |
| DOIs | |
| State | Published - Jun 17 2015 |
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Dimeric organization of blood coagulation factor VIII bound to lipid nanotubes. / Dalm, Daniela; Galaz-Montoya, Jesus G.; Miller, Jaimy L.; Grushin, Kirill; Villalobos, Alex; Koyfman, Alexey Y.; Schmid, Michael F.; Stoilova-McPhie, Svetla.
In: Scientific Reports, Vol. 5, 11212, 17.06.2015.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Dimeric organization of blood coagulation factor VIII bound to lipid nanotubes
AU - Dalm, Daniela
AU - Galaz-Montoya, Jesus G.
AU - Miller, Jaimy L.
AU - Grushin, Kirill
AU - Villalobos, Alex
AU - Koyfman, Alexey Y.
AU - Schmid, Michael F.
AU - Stoilova-McPhie, Svetla
PY - 2015/6/17
Y1 - 2015/6/17
N2 - Membrane-bound Factor VIII (FVIII) has a critical function in blood coagulation as the pro-cofactor to the serine-protease Factor IXa (FIXa) in the FVIIIa-FIXa complex assembled on the activated platelet membrane. Defects or deficiency of FVIII cause Hemophilia A, a mild to severe bleeding disorder. Despite existing crystal structures for FVIII, its membrane-bound organization has not been resolved. Here we present the dimeric FVIII membrane-bound structure when bound to lipid nanotubes, as determined by cryo-electron microscopy. By combining the structural information obtained from helical reconstruction and single particle subtomogram averaging at intermediate resolution (15-20 Å), we show unambiguously that FVIII forms dimers on lipid nanotubes. We also demonstrate that the organization of the FVIII membrane-bound domains is consistently different from the crystal structure in solution. The presented results are a critical step towards understanding the mechanism of the FVIIIa-FIXa complex assembly on the activated platelet surface in the propagation phase of blood coagulation.
AB - Membrane-bound Factor VIII (FVIII) has a critical function in blood coagulation as the pro-cofactor to the serine-protease Factor IXa (FIXa) in the FVIIIa-FIXa complex assembled on the activated platelet membrane. Defects or deficiency of FVIII cause Hemophilia A, a mild to severe bleeding disorder. Despite existing crystal structures for FVIII, its membrane-bound organization has not been resolved. Here we present the dimeric FVIII membrane-bound structure when bound to lipid nanotubes, as determined by cryo-electron microscopy. By combining the structural information obtained from helical reconstruction and single particle subtomogram averaging at intermediate resolution (15-20 Å), we show unambiguously that FVIII forms dimers on lipid nanotubes. We also demonstrate that the organization of the FVIII membrane-bound domains is consistently different from the crystal structure in solution. The presented results are a critical step towards understanding the mechanism of the FVIIIa-FIXa complex assembly on the activated platelet surface in the propagation phase of blood coagulation.
UR - http://www.scopus.com/inward/record.url?scp=84935897774&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84935897774&partnerID=8YFLogxK
U2 - 10.1038/srep11212
DO - 10.1038/srep11212
M3 - Article
VL - 5
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
M1 - 11212
ER -