title: Exponential size distribution of von Willebrand factor.
abstract: Von Willebrand Factor (VWF) is a multimeric protein crucial for hemostasis. Under shear flow, it acts as a mechanosensor responding with a size-dependent globule-stretch transition to increasing shear rates. Here, we quantify for the first time, to our knowledge, the size distribution of recombinant VWF and VWF-eGFP using a multilateral approach that involves quantitative gel analysis, fluorescence correlation spectroscopy, and total internal reflection fluorescence microscopy. We find an exponentially decaying size distribution of multimers for recombinant VWF as well as for VWF derived from blood samples in accordance with the notion of a step-growth polymerization process during VWF biosynthesis. The distribution is solely described by the extent of polymerization, which was found to be reduced in the case of the pathologically relevant mutant VWF-IIC. The VWF-specific protease ADAMTS13 systematically shifts the VWF size distribution toward smaller sizes. This dynamic evolution is monitored using fluorescence correlation spectroscopy and compared to a computer simulation of a random cleavage process relating ADAMTS13 concentration to the degree of VWF breakdown. Quantitative assessment of VWF size distribution in terms of an exponential might prove to be useful both as a valuable biophysical characterization and as a possible disease indicator for clinical applications.
authors.affiliation: Faculty of Physics and Center for NanoScience, Ludwig Maximilian University, Munich, Germany.
authors.forename: Svenja
authors.initials: S
authors.lastname: Lippok
authors.forename: Tobias
authors.initials: T
authors.lastname: Obser
authors.forename: Jochen P
authors.initials: JP
authors.lastname: Müller
authors.forename: Valentin K
authors.initials: VK
authors.lastname: Stierle
authors.forename: Martin
authors.initials: M
authors.lastname: Benoit
authors.forename: Ulrich
authors.initials: U
authors.lastname: Budde
authors.forename: Reinhard
authors.initials: R
authors.lastname: Schneppenheim
authors.forename: Joachim O
authors.initials: JO
authors.lastname: Rädler
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism; D000071120:ADAMTS13 Protein; D006801:Humans; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D059748:Proteolysis; D013050:Spectrometry, Fluorescence; D014841:von Willebrand Factor / Q000737:chemistry* / Q000378:metabolism
chemical_list: D014841:von Willebrand Factor; D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human