Factor VIII organisation on nanodiscs with different lipid composition

Kirill Grushin; Jaimy Miller; Daniela Dalm; Svetla Stoilova-McPhie

doi:10.1160/TH14-09-0725

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2015; 113(04): 741-749
DOI: 10.1160/TH14-09-0725

Coagulation and Fibrinolysis

Schattauer GmbH

Factor VIII organisation on nanodiscs with different lipid composition

Kirill Grushin

¹Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, Texas, USA

,

Jaimy Miller

¹Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, Texas, USA

,

Daniela Dalm

¹Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, Texas, USA

,

Svetla Stoilova-McPhie

¹Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, Texas, USA

²Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, Texas, USA

› Author Affiliations

Abstract

Summary

Nanodiscs (ND) are lipid bilayer membrane patches held by amphiphilic scaffolding proteins (MSP) of ~10 nm in diameter. Nanodiscs have been developed as lipid nanoplatforms for structural and functional studies of membrane and membrane associated proteins. Their size and monodispersity have rendered them unique for electron microscopy (EM) and single particle analysis studies of proteins and complexes either spanning or associated to the ND membrane. Binding of blood coagulation factors and complexes, such as the Factor VIII (FVIII) and the Factor VIIIa - Factor IXa (intrinsic tenase) complex to the negatively charged activated platelet membrane is required for normal haemostasis. In this study we present our work on optimising ND, specifically designed to bind FVIII at close to physiological conditions. The binding of FVIII to the negatively charged ND rich in phosphatidylserine (PS) was followed by electron microscopy at three different PS compositions and two different membrane scaffolding protein (MSP1D1) to lipid ratios. Our results show that the ND with highest PS content (80 %) and lowest MSP1D1 to lipid ratio (1:47) are the most suitable for structure determination of the membrane-bound FVIII by single particle EM. Our preliminary FVIII 3D reconstruction as bound to PS containing ND demonstrates the suitability of the optimised ND for structural studies by EM. Further assembly of the activated FVIII form (FVIIIa) and the whole FVIIIa-FIXa complex on ND, followed by EM and single particle reconstruction will help to identify the protein-protein and protein-membrane interfaces critical for the intrinsic tenase complex assembly and function.

Keywords

Coagulation factors - imaging - phospholipids - electron microscopy - protein structure / folding

Full Text

References

References
1 Fay PJ. Activation of factor VIII and mechanisms of cofactor action. Blood Rev 2004; 18: 1-15.
2 Fay PJ. Factor VIII structure and function. Intern J Hematol 2006; 83: 103-108.
3 Pittman DD, Kaufman RJ. Proteolytic requirements for thrombin activation of anti-haemophilic factor (factor VIII). Proc Natl Acad Sci USA 1988; 85: 2429-2433.
4 Fay PJ, Smudzin TM. Characterisation of the interaction between the A2 subunit and A1/A3-C1-C2 dimer in human factor VIIIa. J Biol Chem 1992; 267: 13246-13250.
5 Wakabayashi H, Fay PJ. Identification of residues contributing to A2 domaindependent structural stability in factor VIII and factor VIIIa. J Biol Chem 2008; 283: 11645-11651.
6 Shen BW, Spiegel PC, Chang CH. et al. The tertiary structure and domain organisation of coagulation factor VIII. Blood 2008; 111: 1240-1247.
7 Derrick TS, Kashi RS, Durrani M. et al. Effect of metal cations on the conformation and inactivation of recombinant human factor VIII. J Pharm Sci 2004; 93: 2549-2557.
8 Lollar P, Parker CG. Subunit structure of thrombin-activated porcine factor VIII. Biochemistry 1989; 28: 666-674.
9 Fay PJ, Haidaris PJ, Smudzin TM. Human factor VIIIa subunit structure. Reconstruction of factor VIIIa from the isolated A1/A3-C1-C2 dimer and A2 subunit. J Biol Chem 1991; 266: 8957-8962.
10 Duffy EJ, Parker ET, Mutucumarana VP. et al. Binding of factor VIIIa and factor VIII to factor IXa on phospholipid vesicles. J Biol Chem 1992; 267: 17006-17011.
11 Pratt KP, Shen BW, Takeshima K. et al. Structure of the C2 domain of human factor VIII at 1.5 A resolution. Nature 1999; 402: 439-442.
12 Lenting PJ, van Mourik JA, Mertens K. The life cycle of coagulation factor VIII in view of its structure and function. Blood 1998; 92: 3983-3996.
13 Hsu TC, Pratt KP, Thompson AR. The factor VIII C1 domain contributes to platelet binding. Blood 2008; 111: 200-208.
14 Ahmad SS, Walsh PN. Coordinate binding studies of the substrate (factor X) with the cofactor (factor VIII) in the assembly of the factor X activating complex on the activated platelet surface. Biochemistry 2002; 41: 11269-11276.
15 Furie B, Furie BC. Mechanisms of thrombus formation. N Engl J Med 2008; 359: 938-949.
16 Pipe SW. Haemophilia: new protein therapeutics. Hematology Am Soc Hematol Educ Program 2010; 2010: 203-209.
17 Shaw AW, Pureza VS, Sligar SG. et al. The local phospholipid environment modulates the activation of blood clotting. J Biol Chem 2007; 282: 6556-6563.
18 Ruprecht J, Nield J. Determining the structure of biological macromolecules by transmission electron microscopy, single particle analysis and 3D reconstruction. Progr Biophys Mol Biol 2001; 75: 121-164.
19 Grushin K, Miller J, Dalm D. et al. Lack of recombinant factor VIII B-domain induces phospholipid vesicle aggregation: implications for the immunogenicity of factor VIII. Haemophilia 2014; 20: 723-731.
20 Henderson R. The potential and limitations of neutrons, electrons and X-rays for atomic resolution microscopy of unstained biological molecules. Quart Rev Biophys 1995; 28: 171-193.
21 Frederik PM, Hubert DHW. Cryoelectron microscopy of liposomes. Methods Enzymol 2005; 391: 431-448.
22 Katayama H, Wang J, Tama F. et al. Three-dimensional structure of the anthrax toxin pore inserted into lipid nanodiscs and lipid vesicles. Proc Natl Acad Sci USA 2010; 107: 3453-3457.
23 Akkaladevi N, Hinton-Chollet L, Katayama H. et al. Assembly of anthrax toxin pore: lethal-factor complexes into lipid nanodiscs. Protein Sci 2013; 22: 492-501.
24 Gatti L, Mannucci PM. Use of porcine factor VIII in the management of seventeen patients with factor VIII antibodies. Thromb Haemost 1984; 51: 379-384.
25 Hay CR, Lozier JN, Lee CA. et al. Safety profile of porcine factor VIII and its use as hospital and home-therapy for patients with haaemophilia-A and inhibitors: the results of an international survey. Thromb Haemost 1996; 75: 25-29.
26 Lollar P, Parker ET. Structural basis for the decreased procoagulant activity of human factor VIII compared to the porcine homolog. J Biol Chem 1991; 266: 12481-12486.
27 Lollar P, Parker ET, Fay PJ. Coagulant properties of hybrid human/porcine factor VIII molecules. J Biol Chem 1992; 267: 23652-23657.
28 Doering CB, Healey JF, Parker ET. et al. High level expression of recombinant porcine coagulation factor VIII. J Biol Chem 2002; 277: 38345-38349.
29 Miller J, Dalm D, Koyfman AY. et al. Helical organisation of blood coagulation factor VIII on lipid nanotubes. J Vis Exp 2014. Epub ahead of print.
30 Stoilova-McPhie S, Grushin K, Dalm D. et al. Lipid nanotechnologies for structural studies of membrane-associated proteins. Proteins 2014; 82: 2902-2909.
31 Ritchie TK, Grinkova YV, Bayburt TH. et al. Chapter 11 - Reconstitution of membrane proteins in phospholipid bilayer nanodiscs. Methods Enzymol 2009; 464: 211-231.
32 Pace CN, Vajdos F, Fee L. et al. How to measure and predict the molar absorption coefficient of a protein. Protein Sci 1995; 04: 2411-2423.
33 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685.
34 Tang G, Peng L, Baldwin PR. et al. EMAN2: an extensible image processing suite for electron microscopy. J Struct Biol 2007; 157: 38-46.
35 Parmenter CDJ, Stoilova-McPhie S. Binding of recombinant human coagulation factor VIII to lipid nanotubes. FEBS Lett 2008; 582: 1657-1660.
36 Blanchette CD, Law R, Benner WH. et al. Quantifying size distributions of nanolipoprotein particles with single-particle analysis and molecular dynamic simulations. J Lipid Res 2008; 49: 1420-1430.
37 Denisov IG, Grinkova YV, Lazarides aa. et al. Directed self-assembly of monodisperse phospholipid bilayer Nanodiscs with controlled size. J Am Chem Soc 2004; 126: 3477-3487.
38 Stoilova-McPhie S, Lynch GC, Ludtke S. et al. Domain organisation of membrane-bound factor VIII. Biopolymers 2013; 99: 448-459.
39 Morrissey JH, Pureza V, Davis-Harrison RL. et al. Blood clotting reactions on nanoscale phospholipid bilayers. Thromb Res 2008; 122 Suppl S23-S26.
40 Grinkova YV, Denisov IG, Sligar SG. Engineering extended membrane scaffold proteins for self-assembly of soluble nanoscale lipid bilayers. Protein engineering design selection PEDS 2010; 23: 843-848.
41 Biró E, Akkerman JWN, Hoek FJ. et al. The phospholipid composition and cholesterol content of platelet-derived microparticles: a comparison with platelet membrane fractions. J Thromb Haemost 2005; 03: 2754-2763.
42 Demel Ra, Paltauf F, Hauser H. Monolayer characteristics and thermal behavior of natural and synthetic phosphatidylserines. Biochemistry 1987; 26: 8659-8665.
43 Smaby JM, Kulkarni VS, Momsen M. et al. The interfacial elastic packing interactions of galactosylceramides, sphingomyelins, and phosphatidylcholines. Biophys J 1996; 70: 868-877.
44 Stoylova SS, Lenting PJ, Kemball-Cook G. et al. Electron crystallography of human blood coagulation factor VIII bound to phospholipid monolayers. J Biol Chem 1999; 274: 36573-36578.
45 Stoilova-McPhie S, Villoutreix BO, Mertens K. et al. 3-Dimensional structure of membrane-bound coagulation factor VIII: modeling of the factor VIII heterodimer within a 3-dimensional density map derived by electron crystallography. Blood 2002; 99: 1215-1223.
46 Stoylova S, Mann KG, Brisson A. Structure of membrane-bound human factor Va. FEBS Lett 1994; 351: 330-334.
47 Stoylova S, Gray E, Barrowcliffe TW. et al. Structural determination of lipidbound human blood coagulation factor IX. Biochim Biophys Acta 1998; 1383: 175-178.

Supplementary Material

Online Supplementary Material (PDF)