Dimeric organization of blood coagulation factor VIII bound to lipid nanotubes

Daniela Dalm; Jesus G. Galaz-Montoya; Jaimy L. Miller; Kirill Grushin; Alex Villalobos; Alexey Y. Koyfman; Michael F. Schmid; Svetla Stoilova-McPhie

doi:10.1038/srep11212

Dimeric organization of blood coagulation factor VIII bound to lipid nanotubes

Daniela Dalm, Jesus G. Galaz-Montoya, Jaimy L. Miller, Kirill Grushin, Alex Villalobos, Alexey Y. Koyfman, Michael F. Schmid, Svetla Stoilova-McPhie

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

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	https://doi.org/10.1038/srep11212
State	Published - Jun 17 2015
Externally published	Yes

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@article{b88fe2f1124c45e98ae3dc08ccfe016c,

title = "Dimeric organization of blood coagulation factor VIII bound to lipid nanotubes",

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 {\AA}), 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.",

author = "Daniela Dalm and Galaz-Montoya, {Jesus G.} and Miller, {Jaimy L.} and Kirill Grushin and Alex Villalobos and Koyfman, {Alexey Y.} and Schmid, {Michael F.} and Svetla Stoilova-McPhie",

note = "Funding Information: This work was supported by a National Scientist Development grant from the American Heart Association: 10SDG3500034 and UTMB start up funds to SSM. The authors acknowledge the cryo-EM and computing facilities at the Sealy Center for Structural Biology (SCSB) at UTMB (Galveston) and the director of the SCSB, Dr. B.M. Pettitt for continuous support, constructive critiques and encouragement. SSM also acknowledges the National Center for Molecular Imaging at Baylor College of Medicine (Houston) supported by NIH grants R01GM080139 and P41GM103832 to Dr. Steven Ludtke and Dr. Wah Chiu. SSM acknowledges Dr. Edward Egelman from the University of Virginia (Charlottesville) and Dr. Pete Lollar and his team at Emory Hospital (Atlanta, Georgia) for support throughout the project.",

year = "2015",

month = jun,

day = "17",

doi = "10.1038/srep11212",

language = "English (US)",

volume = "5",

journal = "Scientific reports",

issn = "2045-2322",

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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

N1 - Funding Information: This work was supported by a National Scientist Development grant from the American Heart Association: 10SDG3500034 and UTMB start up funds to SSM. The authors acknowledge the cryo-EM and computing facilities at the Sealy Center for Structural Biology (SCSB) at UTMB (Galveston) and the director of the SCSB, Dr. B.M. Pettitt for continuous support, constructive critiques and encouragement. SSM also acknowledges the National Center for Molecular Imaging at Baylor College of Medicine (Houston) supported by NIH grants R01GM080139 and P41GM103832 to Dr. Steven Ludtke and Dr. Wah Chiu. SSM acknowledges Dr. Edward Egelman from the University of Virginia (Charlottesville) and Dr. Pete Lollar and his team at Emory Hospital (Atlanta, Georgia) for support throughout the project.

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.

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Dimeric organization of blood coagulation factor VIII bound to lipid nanotubes

Abstract

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