Structural basis of membrane binding by Gla domains of vitamin K–dependent proteins

  title={Structural basis of membrane binding by Gla domains of vitamin K–dependent proteins},
  author={Mingdong Huang and Alan C. Rigby and Xavier Morelli and Marianne A. Grant and Guiqing Huang and Bruce Furie and Barbara A. Seaton and Barbara C. Furie},
  journal={Nature Structural Biology},
In a calcium-dependent interaction critical for blood coagulation, vitamin K–dependent blood coagulation proteins bind cell membranes containing phosphatidylserine via γ-carboxyglutamic acid–rich (Gla) domains. Gla domain–mediated protein-membrane interaction is required for generation of thrombin, the terminal enzyme in the coagulation cascade, on a physiologic time scale. We determined by X-ray crystallography and NMR spectroscopy the lysophosphatidylserine-binding site in the bovine… 

Computational study of coagulation factor VIIa’s affinity for phospholipid membranes

The MD simulation approach provides a structural and dynamic support to the role of P10Q and K32E mutations in the improvement of the membrane contact and a ranking of the rupture force that can be useful in the interpretation of the PS interaction with Gla domains is exhibited.

Crystal Structure of the Calcium-stabilized Human Factor IX Gla Domain Bound to a Conformation-specific Anti-factor IX Antibody*

The fully carboxylated Gla domain of Factor IX is prepared by solid phase peptide synthesis and crystallized Factor IX-(1-47) in complex with Fab fragments of the 10C12 antibody and it is concluded that this antibody is directed at the membrane binding site in the ω loop offactor IX and blocks Factor IX function by inhibiting its interaction with membranes.

Selective modulation of protein C affinity for EPCR and phospholipids by Gla domain mutation

Protein’C binding to sEPCR and phospholipids is broadly dependent on correct Gla domain folding, but can be selectively influenced by judicious mutation.

Molecular Determinants of Phospholipid Synergy in Blood Clotting*

It is proposed that phosphatidylcholine and sphingomyelin (the major external phospholipids of healthy cells) are anticoagulant primarily because their bulky choline headgroups sterically hinder access to their phosphates.

Interaction of prothrombin with a phospholipid surface: evidence for a membrane-induced conformational change

Data suggest that, on binding to a membrane surface, prothrombin undergoes a conformational change to a form which binds more tightly to the membrane.

Lipid specificity of the membrane binding domain of coagulation factor X

Molecular dynamics simulations were used to develop the first membrane‐bound model of FX‐GLA, and potential PS‐specific binding sites identified.

Tissue factor/factor VIIa complex: role of the membrane surface.

Multifunctional Specificity of the Protein C/Activated Protein C Gla Domain*

Results reveal the specific Gla domain residues responsible for mediating protein C/APC molecular recognition with both its cofactor and receptor and further illustrate the multifunctional potential of Gla domains.

The gamma-carboxyglutamic acid domain of anticoagulant protein S is involved in activated protein C cofactor activity, independently of phospholipid binding.

By characterizing PS mutants in which PS face 1 and PS face 2 were individually replaced by the corresponding prothrombin faces, it is found that face 1 was necessary for efficient phospholipid binding but that face 2 residues were not strictly required for phospholIPid binding and were involved in the interaction with APC.



Identification of the Phospholipid Binding Site in the Vitamin K-dependent Blood Coagulation Protein Factor IX*

Results indicate that the NH2 terminus of the Gla domain, specifically including leucine 6 and phenylalanine 9 in the hydrophobic patch, is the contact surface on Factor IX that interacts with the phospholipid bilayer.

Vitamin K-dependent proteins.

Structure of the Ca2+-free GLA domain sheds light on membrane binding of blood coagulation proteins

It is proposed that the Ca2+-induced exposure of hydrophobic side chains is crucial for membrane binding of Gla-containing coagulation proteins.

Crystal structure of an anticoagulant protein in complex with the Gla domain of factor X

The crystal structure of X-bp in complex with the Gla domain peptide of factor X at 2.3-Å resolution showed that the anticoagulation is based on the fact that two patches of the GlA domain essential for membrane binding are buried in the complex formation.

Contributions of Gla and EGF-like domains to the function of vitamin K-dependent coagulation factors.

  • J. Stenflo
  • Biology
    Critical reviews in eukaryotic gene expression
  • 1999
The N-terminal noncatalytic Gla and EGF-like domains that provide the coagulation serine proteases with unique properties, such as affinity for certain biological membranes, and also mediate protein-protein interactions, are the subject of this review.

Comparison of naturally occurring vitamin K-dependent proteins: correlation of amino acid sequences and membrane binding properties suggests a membrane contact site.

Comparison of naturally occurring vitamin K-dependent proteins has provided possible bases for divergent membrane binding and suggested future approaches to determine biological function.

Hydrophobic amino acid residues of human anticoagulation protein C that contribute to its functional binding to phospholipid Vesicles.

It is proposed that the PL-dependent activity losses of these mutants are related to their suboptimal binding to PL or to their misorientation on the PL surface leading to poor alignment of the active sites of the r-APC mutants with the complementary cleavage sites on fVIII/fVIIIa and fV/fVa.

Ca2+ bridges the C2 membrane‐binding domain of protein kinase Cα directly to phosphatidylserine

Three‐dimensional structures of the Ca2+‐bound forms of the PKCα‐C2 domain both in the absence and presence of 1,2‐dicaproyl‐sn‐phosphatidyl‐L‐serine have been determined and suggest a membrane binding mechanism of the C2 domain in which calcium ions directly mediate the phosphate recognition while the calcium binding region 3 might penetrate into the phospholipid bilayer.

The Ca2+ ion and membrane binding structure of the Gla domain of Ca-prothrombin fragment 1.

The structure of Ca-prothrombin fragment 1 has been solved and refined at 2.2-A resolution by X-ray crystallographic methods and Electrostatic potential calculations indicate that the Gla domain is very electronegative.

The Gla Domain of Human Prothrombin Has a Binding Site for Factor Va*

Results indicate that the N-terminal Gla domain of human prothrombin is a functional unit that has a binding site for factor Va, which is important for interaction of the substrate with the pro thirdrombinase complex.