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

Huang, Mingdong; Rigby, Alan C.; Morelli, Xavier; Grant, Marianne A.; Huang, Guanhao; Furie, Bruce; Seaton, Barbara A.; Furie, Barbara C.

doi:10.1038/nsb971

Cited by 214 publications

(237 citation statements)

References 43 publications

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“…(Gla-domaincontaining proteins are prominent components of the blood coagulation cascade.) These Gla domains bind the phospholipid phosphatidylserine 13 , and this is an important feature of the in vivo function of GAS6 and protein S [14][15][16][17] . The Gla domain is followed by four epidermal growth factor (EGF)-like modules, and then by two tandem laminin G domains that are related to those of the sex hormone binding globulin (SHBG) (FIG.…”

Section: Tam Receptors and Ligandsmentioning

confidence: 99%

Immunobiology of the TAM receptors

2008

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Recent studies have revealed that the TAM receptor protein tyrosine kinases -TYRO3, AXL and MER -have pivotal roles in innate immunity. They inhibit inflammation in dendritic cells and macrophages, promote the phagocytosis of apoptotic cells and membranous organelles, and stimulate the maturation of natural killer cells. Each of these phenomena may depend on a cooperative interaction between TAM receptor and cytokine receptor signalling systems. Although its importance was previously unrecognized, TAM signalling promises to have an increasingly prominent role in studies of innate immune regulation.Receptor protein tyrosine kinases (PTKs) are cell-surface transmembrane receptors that contain a regulated PTK activity within their cytoplasmic domains. They function as sensors for extracellular ligands, the binding of which triggers receptor dimerization and activation of the receptor's kinase. This leads to the recruitment, phosphorylation and activation of multiple downstream signalling proteins, which ultimately change the physiology of cells. Although there are only 58 receptor PTK genes in the human genome 1 (see the human kinome website), the signal transduction cascades initiated by receptor PTK activation control diverse cellular processes -from cell differentiation to cell death. Well-known receptor PTK subfamilies include the ERBB receptors, which have essential roles in cardiac and neural development and the progression of some forms of breast cancer 2 ; and the ephrin receptors, which are required for tissue morphogenesis and the patterning of neuronal connections in the developing brain 3 .The focus of this Review, the TAM group, was among the last receptor PTK subfamilies to be identified, and the biological roles of its three members -TYRO3, AXL and MERremained uncharacterized for several years. Largely through the analysis of engineered lossof-function mutants in mice, these roles have become increasingly apparent. They reflect a specific requirement for TAM signalling in settings in which fully differentiated cells, tissues and organs must be maintained in the face of continuous challenge, turnover and renewal. In humans, ongoing homeostatic regulation of this sort must be carried out, frequently on a daily basis, for decades. Although an essential role for TAM regulation of tissue homeostasis is evident in the adult nervous, reproductive and vascular systems, it is in In this Review, we highlight the central roles that TAM signalling has in the intrinsic inhibition of the inflammatory response to pathogens by dendritic cells (DCs) and macrophages; during phagocytosis of apoptotic cells by these same cells; and in the maturation and killing activity of natural killer (NK) cells. We also discuss how, in many or all of these settings, TAM receptors depend on and interact with cytokine receptors. TAM receptors and ligandsThe three TAM receptors, TYRO3, AXL and MER, were identified as a distinct receptor PTK subfamily in 1991 (REFS 4,5 ). Subsequent cloning of full-length cDNAs by multiple labo...

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Section: Tam Receptors and Ligandsmentioning

confidence: 99%

Immunobiology of the TAM receptors

2008

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“…Assuming that PS and prothrombin insert into the membrane up to the Ca 2ϩ ions (insertion of the N-terminal -loop into the membrane), 42 we focused on differences in the solvent-exposed side chains of the portion encompassing residues 10 to 45. The main differences between the PS and prothrombin Gla domains involved 2 distinct clusters of solvent-exposed amino acid residues located on 2 opposite faces.…”

Section: Structural Analysismentioning

confidence: 99%

“…Although Gln10 was part of face 1 of the PS Gla domain, it belonged to the PS loop, thought to be a strong determinant of phospholipid binding in vitamin K-dependent proteins. 42 We did not, therefore, mutate this residue in F1 FII -Gla PS -PS because this might have impaired phospholipid binding. PS has a potentially The capacity of each PS species to act as an APC cofactor in FVa inactivation was quantified by determining the rate of FXacatalyzed prothrombin activation in a reaction mixture containing 2 nM FV/FVa, 0.1 nM FXa, 100 nM prothrombin, and 10 M PC/PS vesicles, as described in "Materials and methods."…”

Section: Probing the Involvement Of Faces 1 And 2 Of The Ps Gla Domaimentioning

confidence: 99%

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

Saller

Villoutreix

Amelot

et al. 2005

Blood

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We expressed 2 chimeras between human protein S (PS) and human prothrombin (FII) in which the prothrombin ␥-carboxyglutamic acid (Gla) domain replaced the PS Gla domain in native PS (Gla FII -PS) or in PS deleted of the thrombinsensitive region (TSR) (Gla FII -⌬TSR-PS). Neither PS/FII chimera had activated protein C (APC) cofactor activity in plasma clotting assays or purified systems, but both bound efficiently to phospholipids. This pointed to a direct involvement of the PS Gla domain in APC cofactor activity through molecular interaction with APC. Using computational methods, we identified 2 opposite faces of solventexposed residues on the PS Gla domain (designated faces 1 and 2) as potentially involved in this interaction. Their importance was supported by functional characterization of a PS mutant in which the face 1 and face 2 PS residues were reintroduced into Gla FII -PS, leading to significant APC cofactor activity, likely through restored interaction with APC. Furthermore, by characterizing PS mutants in which PS face 1 and PS face 2 were individually replaced by the corresponding prothrombin faces, we 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. IntroductionProtein S (PS) is a vitamin K-dependent protein involved in regulating the anticoagulant activity of activated protein C (APC). In purified systems, PS functions as a nonenzymatic cofactor for APC in proteolytic inactivation of activated factor V (FVa) and activated factor VIII (FVIIIa), 1 reflecting the ability of PS to interact with APC. However, these effects are relatively weak and are inconsistent with the major physiologic role of PS. Indeed, the importance of PS as a natural anticoagulant is clearly demonstrated by the occurrence of severe thrombotic complications in infants with homozygous hereditary PS deficiency and by a mild thrombotic tendency in heterozygous subjects. 2 This suggests that additional components may determine the expression of the APC cofactor activity of PS in plasma. Thus, activated factor X (FXa) and activated factor IX (FIXa) have been shown to protect FVa and FVIIIa from inactivation by APC, and, in this system, PS is able to abolish these protective effects. [3][4][5] Another determinant is FV, which acts in synergy with PS in FVIIIa inactivation by APC in purified systems. 6 PS also demonstrates APC-independent anticoagulant activity by inhibiting prothrombinase and tenase activities, 7,8 and this may contribute to the overall anticoagulant activity of PS in plasma.PS is a single-chain, 635-amino acid glycoprotein composed of an N-terminal domain rich in ␥-carboxyglutamic acid (Gla domain; amino acids 1-45), a thrombin-sensitive region (TSR; amino acids 46-74), 4 epidermal growth factor (EGF)-like domains (amino acids 75-242), 9 and a large C-terminal sex hormone-binding globulin (SHBG)-like region (amino acids 243-635). 10 The SHBGlike domain of PS binds tightly to C4b...

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“…Somewhat surprisingly, despite the fact that OCN is a secreted protein, these investigators did not need to permeabilize the cells in order to detect cells producing OCN. The reasons why this was possible are somewhat unclear, but may have to do with the fact that OCN does possess Gla residues [5], which may allow for at least temporary anchoring of the protein to the cell membrane as the protein is secreted, similar to the mechanisms by which Gla residues on clotting factors allow attachment of these proteins to cell membranes [6]. Alternatively, OCN producing cells may also possess an OCN receptor [7,8], and as the protein is secreted, it may bind to this cell surface receptor.…”

Section: Introductionmentioning

confidence: 99%

Characterization of circulating osteoblast lineage cells in humans

Eghbali‐Fatourechi

Mödder

Charatcharoenwitthaya

et al. 2007

Bone

123

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We recently identified circulating osteoblastic cells using antibodies to osteocalcin (OCN) or alkaline phosphatase (AP). We now provide a more detailed characterization of these cells. Specifically, we demonstrate that 46% of OCN positive (OCN pos ) cells express AP, and 37% also express the hematopoietic/endothelial marker, CD34. Using two different anti-OCN antibodies and forward/side light scatter characteristics by flow cytometry, we find that OCN pos cells consist of two distinct populations: one population exhibits low forward/side scatter, consistent with a small cell phenotype with low granularity, and a second population has higher forward/side scatter (larger and more granular cell). The smaller, low granularity population also co-expresses CD34, whereas the larger, more granular cells are CD34 negative. Using samples from 26 male subjects aged 28 to 68 years, we demonstrate that the concentration of circulating OCN pos cells increases as a function of age (R = 0.59, P = 0.002). By contrast, CD34 pos cells tend to decrease with age (R = −0.31, P = 0.18); as a consequence, the ratio of OCN pos :CD34 pos cells also increases significantly with age (R = 0.54, P = 0.022). These findings suggest significant overlap between circulating cells expressing OCN and those expressing the hematopoietic/endothelial marker, CD34. Further studies are needed to define the precise role of circulating OCN pos cells not only in bone remodeling but rather also potentially in the response to vascular injury.

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

Cited by 214 publications

References 43 publications

Immunobiology of the TAM receptors

Immunobiology of the TAM receptors

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

Characterization of circulating osteoblast lineage cells in humans

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