title: An experimental model to study the in vivo survival of von Willebrand factor. Basic aspects and application to the R1205H mutation.
abstract: To explore the molecular basis of von Willebrand factor (VWF) clearance, an experimental model employing VWF-deficient mice was developed. Biodistribution was examined by the injection of radiolabeled VWF, which was primarily directed to the liver with minor amounts in other organs. Disappearance of VWF from plasma was characterized by a rapid initial phase (t((1/2))alpha = 13 min) and a slow secondary phase (t((1/2))beta = 3 h), with a mean residence time (MRT) of 2.8 h. A similar clearance was observed for VWF consisting of only high or low molecular weight multimers, indicating that, in our experimental model, clearance is independent of multimeric distribution. This allowed us to compare the survival of full-length VWF to truncated variants. Deletion of both the amino-terminal D'-D3 and carboxyl-terminal D4-CK domains resulted in a fragment with a similar clearance to wild-type VWF. Deletion of only the D'-D3 region was associated with an almost 2-fold lower recovery and increased clearance (MRT = 1.6 h), whereas deletion of only the D4-CK region resulted in a significantly reduced clearance (MRT = 4.5 h, p < 0.02). These results point to a role of the D'-D3 region in preventing clearance of VWF. Furthermore, replacement of D3 domain residue Arg-1205 by His resulted in a markedly increased clearance (MRT = 0.3 h; p = 0.004). Therefore, this mutation seems to abrogate the protective effect of the D'-D3 region. In vitro analysis of this mutant also revealed a 2-fold reduced affinity for VWF propeptide at low pH, showing that mutation of Arg-1205 results not only in an increased clearance rate but is also associated with an impaired pH-dependent interaction with VWF propeptide.
authors: Laboratory for Thrombosis and Haemostasis, Department of Haematology (G.03.647), University Medical Center Utrecht, Heidelberglaan 100, 3465 CX Utrecht, The Netherlands. p.j.lenting@lab.azu.nl
authors: Peter J
authors: PJ
authors: Lenting
authors: Erik
authors: E
authors: Westein
authors: Virginie
authors: V
authors: Terraube
authors: Anne-Sophie
authors: AS
authors: Ribba
authors: Eric G
authors: EG
authors: Huizinga
authors: Dominique
authors: D
authors: Meyer
authors: Philip G
authors: PG
authors: de Groot
authors: Cécile V
authors: CV
authors: Denis
mesh_terms: D000818:Animals; D001120:Arginine / Q000737:chemistry; D002460:Cell Line; D003094:Collagen / Q000737:chemistry; D006224:Cricetinae; D017353:Gene Deletion; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000737:chemistry; D010980:Platelet Membrane Glycoproteins; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D020349:Surface Plasmon Resonance; D013997:Time Factors; D014018:Tissue Distribution; D014841:von Willebrand Factor / Q000737:chemistry* / Q000235:genetics / Q000493:pharmacokinetics
chemical_list: D010455:Peptides; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D011994:Recombinant Proteins; C032193:glycoprotein receptor GPIb-IX; D014841:von Willebrand Factor; D006639:Histidine; D003094:Collagen; D001120:Arginine