Subscribe to RSS
DOI: 10.1160/TH10-02-0081
Comparison of plasma-derived and recombinant von Willebrand factor by atomic force microscopy
Publication History
Received:
01 February 2010
Accepted after minor revision:
27 April 2010
Publication Date:
23 November 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/201003/lookinside/thumbnails/10-1160-th10-02-0081-1.jpg)
Summary
Human plasma protein von Willebrand factor (VWF) is composed of a series of multimers with molecular weights ranging from 600 to 20,000 kDa or even more. Plasma-derived VWF (pdVWF) and recombinant VWF (rVWF) differ in that the ultra-large molecular weight multimer portion present in rVWF is usually missing in pdVWF due to partial cleavage of VWF by the plasma protease ADAMTS13. Here, tapping mode atomic force microscopy (TM-AFM) was used to visualise the shape and size of rVWF and pdVWF. The morphology of the variants of VWF was comparable, containing both globular and stretched domains. Mean chain lengths of the filaments and diameters of the core globular domains were determined and analysed on a statistical basis. About 72% of the pdVWF molecules and 70% of the rVWF molecules were 100–300 nm long. The portion of very long molecules (>300 nm) was only slightly greater in rVWF than in pdVWF (20% vs. 18%). The diameters of the globular core structures were in the range of 12 to 30 nm for both types of VWF. Inspection of a purified rVWF dimer revealed a similar range for the globular domain (14–32 nm). Finally, we demonstrate a dramatic conformational change for rVWF upon exposure to high shear stress, as has been reported for pdVWF. Our TM-AFM data show that the overall structure of rVWF is similar to that of pdVWF and that rVWF will extend its conformation under shear stress, which is required to exert its function in primary haemostasis.
-
References
- 1 Sadler JE. Biochemistry and genetics of von Willebrand factor. Annu Rev Biochem 1998; 67: 395-424.
- 2 Ruggeri ZM. Von Willebrand factor: looking back and looking forward. Thromb Haemost 2007; 98: 55-62.
- 3 Sadler JE, Mannucci PM, Berntorp E. et al. Impact, diagnosis and treatment of von Willebrand disease. Thromb Haemost 2000; 84: 160-174.
- 4 Levy GG, Nichols WC, Lian EC. et al. Mutations in a member of the ADAMTS gene family cause thrombotic thrombocytopenic purpura. Nature 2001; 413: 488-494.
- 5 Plaimauer B, Zimmermann K, Volkel D. et al. Cloning, expression, and functional characterization of the von Willebrand factor-cleaving protease (ADAMTS13). Blood 2002; 100: 3626-3632.
- 6 Sadler JE. Von Willebrand factor, ADAMTS13, and thrombotic thrombocytopenic purpura. Blood 2008; 112: 11-18.
- 7 Tsai HM. Physiologic cleavage of von Willebrand factor by a plasma protease is dependent on its conformation and requires calcium ion. Blood 1996; 87: 4235-4244.
- 8 Fischer BE, Schlokat U, Reiter M. et al. Biochemical and functional characterization of recombinant von Willebrand factor produced on a large scale. Cell Mol Life Sci 1997; 53: 943-950.
- 9 Turecek PL, Mitterer A, Matthiessen HP. et al. Biochemical and functional characterization of a rVWF drug candidate produced under serum-free conditions. J Thromb Haemost 2007; 5 (Suppl. 02) P-W-196.
- 10 Varadi K, Rottensteiner H, Vejda S. et al. Species-dependent variability of ADAMTS13-mediated proteolysis of human recombinant von Willebrand factor. J Thromb Haemost 2009; 7: 1134-1142.
- 11 Fowler WE, Fretto LJ, Hamilton KK. et al. Substructure of human von Willebrand factor. J Clin Invest 1985; 76: 1491-1500.
- 12 Ohmori K, Fretto LJ, Harrison RL. et al. Electron microscopy of human factor VIII/Von Willebrand glycoprotein: effect of reducing reagents on structure and function. J Cell Biol 1982; 95: 632-640.
- 13 Fretto LJ, Fowler WE, McCaslin DR. et al. Substructure of human von Willebrand factor. Proteolysis by V8 and characterization of two functional domains. J Biol Chem 1986; 261: 15679-15689.
- 14 Raghavachari M, Tsai H, Kottke-Marchant K. et al. Surface dependent structures of von Willebrand factor observed by AFM under aqueous conditions. Colloids Surf B: Biointerfaces 2000; 19: 315-324.
- 15 Siedlecki CA, Lestini BJ, Kottke-Marchant KK. et al. Shear-dependent changes in the three-dimensional structure of human von Willebrand factor. Blood 1996; 88: 2939-2950.
- 16 Marchant RE, Lea AS, Andrade JD. et al. Interactions of von Willebrand factor on mica by Atomic force Microscopy. J Colloid Interface Sci 1992; 148: 261-272.
- 17 Parot P, Dufrene YF, Hinterdorfer P. et al. Past, present and future of atomic force microscopy in life sciences and medicine. J Mol Recognit 2007; 20: 418-431.
- 18 Binnig G, Quate CF, Gerber C. Atomic force microscope. Phys Rev Lett 1986; 56: 930-933.
- 19 Hansma HG, Kim KJ, Laney DE. et al. Properties of biomolecules measured from atomic force microscope images: a review. J Struct Biol 1997; 119: 99-108.
- 20 Turecek PL, Mitterer A, Matthiessen HP. et al. Development of a plasma- and albumin-free recombinant von Willebrand factor. Hamostaseologie 2009; 29 (Suppl. 01) S32-S38.
- 21 Turecek PL, Gritsch H, Pichler L. et al. In vivo characterization of recombinant von Willebrand factor in dogs with von Willebrand disease. Blood 1997; 90: 3555-3567.
- 22 Schneider SW, Nuschele S, Wixforth A. et al. Shear-induced unfolding triggers adhesion of von Willebrand factor fibers. Proc Natl Acad Sci USA 2007; 104: 7899-7903.
- 23 Hoyer LW, Shainoff JR. Factor VIII-related protein circulates in normal human plasma as high molecular weight multimers. Blood 1980; 55: 1056-1059.
- 24 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor VIII/von Willebrand factor. Blood 1981; 57: 1140-1143.
- 25 Fischer BE, Schlokat U, Mitterer A. et al. Structural analysis of recombinant von Willebrand factor produced at industrial scale fermentation of transformed CHO cells co-expressing recombinant furin. FEBS Lett 1995; 375: 259-262.
- 26 Singh I, Shankaran H, Beauharnois ME. et al. Solution structure of human von Willebrand factor studied using small angle neutron scattering. J Biol Chem 2006; 281: 38266-38275.
- 27 Slayter H, Loscalzo J, Bockenstedt P. et al. Native conformation of human von Willebrand protein. Analysis by electron microscopy and quasi-elastic light scattering. J Biol Chem 1985; 260: 8559-8563.
- 28 Schwarz HP, Turecek PL, Pichler L. et al. Recombinant von Willebrand factor. Thromb Haemost 1997; 78: 571-576.
- 29 Yokota H, Sunwoo J, Sarikaya M. et al. Spin-stretching of DNA and protein molecules for detection by fluorescence and atomic force microscopy. Anal Chem 1999; 71: 4418-4422.
- 30 Furlan M, Robles R, Lämmle B. Partial Purification and characterization of a pro-tease from human plasma cleaving von Willebrand factor to fragments produced by in vivo proteolysis. Blood 1996; 87: 4223-4234.
- 31 Wu T, Lin J, Cruz MA. et al. Force-induced cleavage of single VWFA1A2A3 trido-mains by ADAMTS-13. Blood 2010; 115: 370-378.
- 32 Zhang X, Halvorsen K, Zhang CZ. et al. Mechanoenzymatic cleavage of the ultra-large vascular protein von Willebrand factor. Science 2009; 324: 1330-1334.
- 33 Baldauf C, Schneppenheim R, Stacklies W. et al. Shear-induced unfolding activates von Willebrand factor A2 domain for proteolysis. J Thromb Haemost 2009; 7: 2096-2105.