Exponential size distribution of von Willebrand factor
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Exponential size distribution of von Willebrand factor. / Lippok, Svenja; Obser, Tobias; Müller, Jochen P; Stierle, Valentin K; Benoit, Martin; Budde, Ulrich; Schneppenheim, Reinhard; Rädler, Joachim O.
In: BIOPHYS J, Vol. 105, No. 5, 03.09.2013, p. 1208-16.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Exponential size distribution of von Willebrand factor
AU - Lippok, Svenja
AU - Obser, Tobias
AU - Müller, Jochen P
AU - Stierle, Valentin K
AU - Benoit, Martin
AU - Budde, Ulrich
AU - Schneppenheim, Reinhard
AU - Rädler, Joachim O
N1 - Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.
PY - 2013/9/3
Y1 - 2013/9/3
N2 - 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.
AB - 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.
KW - ADAM Proteins
KW - Humans
KW - Protein Multimerization
KW - Protein Structure, Quaternary
KW - Proteolysis
KW - Spectrometry, Fluorescence
KW - von Willebrand Factor
U2 - 10.1016/j.bpj.2013.07.037
DO - 10.1016/j.bpj.2013.07.037
M3 - SCORING: Journal article
C2 - 24010664
VL - 105
SP - 1208
EP - 1216
JO - BIOPHYS J
JF - BIOPHYS J
SN - 0006-3495
IS - 5
ER -
