Structure of membrane‐bound human factor Va

Stoylova, Svetla; Mann, Kenneth G.; Brisson, Alain

doi:10.1016/0014-5793(94)00881-7

Cited by 34 publications

(32 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although a structural rearrangement due to APC inactivation cannot be completely ruled out, several pieces of evidence argue against this possibility. First, reconstructions of factor Va using electron microscopy (EM) depict a molecule extending Ϸ100 Å from the cell membrane (42), and these dimensions correlate well with the more recent 15-Å EM projection structure of factor VIIIa (43). In homology models of factors Va and VIIIa based on these EM data, a variety of domain orientations have been proposed (Fig.…”

Section: Domain Interfacesmentioning

confidence: 73%

The crystal structure of activated protein C-inactivated bovine factor Va: Implications for cofactor function

Adams

Hockin

Mann

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

142

157

View full text Add to dashboard Cite

In vertebrate hemostasis, factor Va serves as the cofactor in the prothrombinase complex that results in a 300,000-fold increase in the rate of thrombin generation compared with factor Xa alone. Structurally, little is known about the mechanism by which factor Va alters catalysis within this complex. Here, we report a crystal structure of protein C inactivated factor Va (A1⅐A3-C1-C2) that depicts a previously uncharacterized domain arrangement. This orientation has implications for binding to membranes essential for function. A high-affinity calcium-binding site and a copperbinding site have both been identified. Surprisingly, neither shows a direct involvement in chain association. This structure represents the largest physiologically relevant fragment of factor Va solved to date and provides a new scaffold for the future generation of models of coagulation cofactors.

show abstract

Section: Domain Interfacesmentioning

confidence: 73%

The crystal structure of activated protein C-inactivated bovine factor Va: Implications for cofactor function

Adams

Hockin

Mann

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

142

157

View full text Add to dashboard Cite

show abstract

“…The protein density in projection of a FVIII molecule is similar to the surface covered by one FVa molecule, defined by the same type of structural study (31). FV is the closest known protein to FVIII by sequence identity, and FVa has the same domain composition (A1-A2/A3-C1-C2) as the FVIII heterodimer lacking the B domain.…”

Section: Resultsmentioning

confidence: 88%

“…4). We propose that the C1 and C2 domains double ␤-barrel structure is oriented perpendicular to the membrane plane (27,31) and overlapped by the A3 and A1 domains, giving this most prominent density peak in the projection map (Figs. 4 and 5).…”

Section: Resultsmentioning

confidence: 99%

“…Previously, the projection structure of FVa by electron crystallography, has been reported (31). It was of interest therefore to compare the two-dimensional maps of FVa and FVIII (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…6, A and B), are based on the known atomic structures of FXa and FIXa from x-ray crystallography (45,46) and the projection structures of FVIII and FVa from EM crystallography (31). The dimensions of the enzyme, FIXa and FXa determine the position of the enzymebinding regions within the FVa and FVIII, respectively.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Electron Crystallography of Human Blood Coagulation Factor VIII Bound to Phospholipid Monolayers

Stoylova

Lenting²,

Kemball‐Cook

et al. 1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

Coagulation factor VIII binds to negatively charged platelets prior to assembly with the serine protease, factor IXa, to form the factor X-activating enzyme (FXase) complex. The macromolecular organization of membrane-bound factor VIII has been studied by electron crystallography for the first time. For this purpose twodimensional crystals of human factor VIII were grown onto phosphatidylserine-containing phospholipid monolayers, under near to physiological conditions (pH and salt concentration). Electron crystallographic analysis revealed that the factor VIII molecules were organized as monomers onto the lipid layer, with unit cell dimensions: a ‫؍‬ 81.5Å, b ‫؍‬ 67.2 Å, ␥ ‫؍‬ 66.5°, P1 symmetry. Based on a homology-derived molecular model of the factor VIII (FVIII) A domains, the FVIII projection structure solved at 15-Å resolution presents the A1, A2, and A3 domain heterotrimer tilted approximately 65°relative to the membrane plane. The A1 domain is projecting on top of the A3, C1, and C2 domains and with the A2 domain protruding partially between A1 and A3. This organization of factor VIII allows the factor IXa protease and epidermal growth factor-like domain binding sites (localized in the A2 and A3 domains, respectively) to be situated at the appropriate position for the binding of factor IXa. The conformation of the lipid-bound FVIII is therefore very close to that for the activated factor VIIIa predicted in the FX-ase complex.Factor VIII (FVIII) 1 is an essential protein in blood coagulation (1). Deficiency in FVIII is responsible for hemophilia A (classic hemophilia), an X-chromosome-linked bleeding disorder (2). During coagulation, FVIII is proteolytically cleaved to an unstable active heterodimeric form (FVIIIa) by trace amounts of thrombin or factor Xa (FXa) (3). FVIIIa functions as a cofactor, responsible for the efficient activation of factor X (FX) by activated factor IX (FIXa) in the presence of negatively charged phospholipids (PL) and Ca 2ϩ ion (4). The assembly of the membrane-bound FX-ase complex (FX/FVIIIa/FIXa) results in the release of FXa, which associates with the cofactor factor Va (FVa) and prothrombin into the membrane-bound prothrombinase complex. The released thrombin further cleaves fibrinogen to insoluble fibrin, which stabilizes the growing clot and arrests bleeding (5).The binding of FVIIIa to FIXa enhances the catalytic reaction by at least 5 orders of magnitude (6). Although the molecular mechanism of this amplifying effect is not yet known, it seems to be strongly related to the formation of the FX-ase complex onto negatively charged membranes: increasing both the collision frequencies by restricting the reactants to a twodimensional surface, and the specificity of the FIXa active site (7,8).Human FVIII is a large plasma glycoprotein synthesized as a single-chain polypeptide with a predicted molecular mass of 264,763 Da, corresponding to the mature 2332 amino acid sequence, determined from translation of the cloned gene. With N-linked carbohydrate chains, the molecular mass ...

show abstract

Domain organization of membrane‐bound factor VIII

et al. 2013

View full text Add to dashboard Cite

Factor VIII (FVIII) is the blood coagulation protein which when defective or deficient causes for hemophilia A, a severe hereditary bleeding disorder. Activated FVIII (FVIIIa) is the cofactor to the serine protease factor IXa (FIXa) within the membrane-bound Tenase complex, responsible for amplifying its proteolytic activity more than 100,000 times, necessary for normal clot formation. FVIII is composed of two noncovalently linked peptide chains: a light chain (LC) holding the membrane interaction sites and a heavy chain (HC) holding the main FIXa interaction sites. The interplay between the light and heavy chains (HCs) in the membrane-bound state is critical for the biological efficiency of FVIII. Here, we present our cryo-electron microscopy (EM) and structure analysis studies of human FVIII-LC, when helically assembled onto negatively charged single lipid bilayer nanotubes. The resolved FVIII-LC membrane-bound structure supports aspects of our previously proposed FVIII structure from membrane-bound two-dimensional (2D) crystals, such as only the C2 domain interacts directly with the membrane. The LC is oriented differently in the FVIII membrane-bound helical and 2D crystal structures based on EM data, and the existing X-ray structures. This flexibility of the FVIII-LC domain organization in different states is discussed in the light of the FVIIIa-FIXa complex assembly and function.

show abstract

Structure of membrane‐bound human factor Va

Cited by 34 publications

References 42 publications

The crystal structure of activated protein C-inactivated bovine factor Va: Implications for cofactor function

The crystal structure of activated protein C-inactivated bovine factor Va: Implications for cofactor function

Electron Crystallography of Human Blood Coagulation Factor VIII Bound to Phospholipid Monolayers

Domain organization of membrane‐bound factor VIII

Contact Info

Product

Resources

About