Four Hydrophobic Amino Acids of the Factor VIII C2 Domain Are Constituents of Both the Membrane-binding and von Willebrand Factor-binding Motifs

Gilbert, Gary E.; Kaufman, Randal J.; Arena, Andrew A.; Miao, Hongzhi; Pipe, Steven W.

doi:10.1074/jbc.m104732200

Cited by 111 publications

(159 citation statements)

References 50 publications

Supporting

Mentioning

148

Contrasting

Unclassified

Order By: Relevance

“…This patch is highly reminiscent of the hydrophobic surface around the spikes observed for both FaV (W26, W27, M83, L79) [10,33] and FaVIII (V2223, M2199, F2200, L2251, L2252) [11,34], which are likely candidates for phospholipid bilayer binding. Membrane involvement seems to be a com- mon feature for several members of the discoidin domain family.…”

Section: Resultsmentioning

confidence: 99%

Effects of pathological mutations on the stability of a conserved amino acid triad in retinoschisin

Fraternali¹,

Cavallo

Musco

2003

FEBS Letters

View full text Add to dashboard Cite

A three-dimensional model has been calculated for the discoidin domain of retinoschisin (RS1), the protein involved in the X-linked juvenile retinoschisis. The model allows for a mapping of the pathological retinoschisis missense mutations and a rationale for the structural e¡ects of an evolutionary conserved surface exposed triad (W122^R200^W163). Molecular dynamics simulations of the triad mutants models, together with ab initio energy calculations of the complexes corresponding to the triad show that the observed pathological mutations sensibly destabilize local interactions and the entire fold. Moreover the presented model reveals evidence of a putative site for membrane association. ß

show abstract

Section: Resultsmentioning

confidence: 99%

Effects of pathological mutations on the stability of a conserved amino acid triad in retinoschisin

Fraternali¹,

Cavallo

Musco

2003

FEBS Letters

View full text Add to dashboard Cite

show abstract

“…A 1.5-Å x-ray crystal structure of the FVIII C2 domain and a mutagenesis study suggested that two hydrophobic loops formed by Met 2199 -Phe 2200 and Leu 2251 -Leu 2255 play an important role (11,12). Two additional loops formed by Trp 2313 -His 2315 and Gln 2222 -Lys 2227 ) were also proposed to be involved in membrane binding based on an electron microscopy study (13).…”

Section: Blood Coagulation Factor VIII (Fviii)mentioning

confidence: 99%

Trp2313-His2315 of Factor VIII C2 Domain Is Involved in Membrane Binding

Liu

Lin

Yuan

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Factor VIII (FVIII) plays a critical role in blood coagulation by forming the tenase complex with factor IXa and calcium ions on a membrane surface containing negatively charged phospholipids. The tenase complex activates factor X during blood coagulation. Blood coagulation factor VIII (FVIII)2 is synthesized as a single polypeptide chain, including a 19-residue signal peptide.The mature FVIII contains 2,332 amino acid residues arranged within five domains organized as A1-A2-B-A3-C1-C2 (1, 2). FVIII circulates in the blood as a heterodimer: the A1, A2, and variable portions of the B domain forming the heavy chain with a molecular weight varying between 90,000 and 200,000; and the A3, C1 and C2 domains forming the light chain with a molecular weight of 80,000. The noncovalent assembly of the heterodimer is facilitated by divalent metal ions, as revealed by recent crystallographic studies (3, 4). In plasma, FVIII circulates as a complex with von Willebrand factor which stabilizes FVIII by preventing its rapid clearance from the blood circulation (5). During blood coagulation, FVIII is cleaved by thrombin at Arg 372 , Arg 740 , and Arg 1689 (6), which converts it to a fully active cofactor, FVIIIa. FVIIIa dissociates from von Willebrand factor and binds to membrane surfaces where it assembles with the serine protease factor IXa. The presence of FVIIIa enhances the V max for factor X activation by FIXa by 200,000-fold (7). This large amplification requires that the FVIII level in blood be controlled. Mutations of FVIII cause hemophilia A, an X-linked bleeding disorder (8). The molecular mechanisms of FVIII binding to membranes are not fully understood.Several lines of evidence suggest that the light chain of FVIII, in particular the C2 domain, is responsible for the specific binding to membrane surfaces (9, 10). A 1.5-Å x-ray crystal structure of the FVIII C2 domain and a mutagenesis study suggested that two hydrophobic loops formed by Met 2199 -Phe 2200 and Leu 2251 -Leu 2255 play an important role (11,12). Two additional loops formed by Trp 2313 -His 2315 and Gln 2222 -Lys 2227 ) were also proposed to be involved in membrane binding based on an electron microscopy study (13).C2 domain membrane-binding sites were proposed as a drug target to regulate the functional concentrations of coagulation factors, FVIII and FV, that contain a C2 domain (14). A number of small organic molecules were identified that disrupt C2 domain membrane anchoring of FVIII (14) and FV (15). One of these inhibitors, 005B10, identified from in silico screening, was found to inhibit FVIII interaction with negatively charged phospholipids (15). In this study, we report the high resolution crystal structure (1.15 Å) of the human FVIII C2 domain in * This work was supported, in whole or in part, by National Institutes of Health Grant HL086584. This work was also supported by grants from the Natural Science Foundation of China (30811130467 and 30625011), the Ministry of Science and Technology (2006AA02A313 and 2007CB914304), the Chinese Academy ...

show abstract

“…21,22 Mutagenesis studies have confirmed the role of these residues in phospholipid binding. 23,24 The homology of the lactadherin C domains with those of factors VIII and V suggests that similar phospholipid-binding properties may exist. Indeed, lactadherin has been found to bind selectively to PS 25 and to use primarily the C2 domain in its lipid binding.…”

Section: Introductionmentioning

confidence: 99%

Lactadherin inhibits enzyme complexes of blood coagulation by competing for phospholipid-binding sites

Shi

Gilbert

2003

Blood

Self Cite

166

151

View full text Add to dashboard Cite

Lactadherin, a glycoprotein of the milk-fat globule membrane, contains tandem C domains with homology to discoidin-type lectins and to membrane-binding domains of blood-clotting factors V and VIII. We asked whether the structural homology confers the capacity to compete for the membrane-binding sites of factor VIII and factor V and to function as an anticoagulant. Our results indicate that lactadherin competes efficiently with factor VIII and factor V for binding sites on synthetic phosphatidylserine-containing membranes with half-maximal displacement at lactadherin concentrations of 1 to 4 nM. Binding competition correlated to functional inhibition of factor VIIIa-factor IXa (factor Xase) enzyme complex. In contrast to annexin V, lactadherin was an efficient inhibitor of the prothrombinase and the factor Xase complexes regardless of the degree of membrane curvature and the phosphatidylserine content. Lactadherin also inhibited the factor VIIa-tissue factor complex efficiently whereas annexin V was less effective. Because the inhibitory concentration of lactadherin was proportional to the phospholipid concentration, and because lactadherin was not an efficient inhibitor in the absence of phospholipid, the major inhibitory effect of lactadherin relates to blocking phospholipid sites rather than forming inhibitory protein-protein complexes. Lactadherin was also an effective inhibitor of a modified whole blood prothrombin time assay in which clotting was initiated by dilute tissue factor; 60 nM lactadherin prolonged the prothrombin time 150% IntroductionLactadherin is a 47 000-Da molecular weight (MW) glycoprotein of milk-fat globules. It has also been known as PAS-6/7, indicating the 2 glycosylation variants, 1 bovine-associated mucoprotein, BA-46, P47, and MFG-E8. 2 Lactadherin has a domain structure of EGF1-EGF2-C1-C2 in which EGF indicates epidermal growth factor homology domains, and the C domains share homology with the discoidin family, including the lipid-binding C domains of blood coagulation factor VIII and factor V. 2 The second EGF domain displays an Arg-Gly-Asp motif, 3 which binds to the ␣ v ␤ 3 and ␣ v ␤ 5 integrins. 1,[4][5][6] The second C domain binds to phospholipids. 6 In milk-fat globules, lactadherin lines the surface of the phospholipid bilayer that surrounds the central triglyceride droplet, apparently stabilizing the bilayer. 7 Lactadherin decreases the symptoms of rotavirus infection in infants, possibly by binding to rotavirus particles via carbohydrate moieties. 8 In tissue sections, lactadherin is found localized on the apical portion of secretory epithelium in the breast. 7 Abundant expression by breast carcinoma tissue makes lactadherin a potential target for antigen-guided radiation therapy. 9 Lactadherin is also found on the apical surface of epithelia in the biliary tree, the pancreas, and sweat glands 7 and is synthesized by aortic medial smooth muscle cells. 10 Function in these tissues remains unknown. Lactadherin has been identified as a zona pellucida-binding protein on the a...

show abstract

Four Hydrophobic Amino Acids of the Factor VIII C2 Domain Are Constituents of Both the Membrane-binding and von Willebrand Factor-binding Motifs

Cited by 111 publications

References 50 publications

Effects of pathological mutations on the stability of a conserved amino acid triad in retinoschisin

Effects of pathological mutations on the stability of a conserved amino acid triad in retinoschisin

Trp2313-His2315 of Factor VIII C2 Domain Is Involved in Membrane Binding

Lactadherin inhibits enzyme complexes of blood coagulation by competing for phospholipid-binding sites

Contact Info

Product

Resources

About