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Efficient Production and Purification of Recombinant Murine Kindlin-3 from Insect Cells for Biophysical Studies
Published on: March 19, 2014
High-molecular-weight kininogen is a binding protein for tissue prokallikrein
1Institute for Biochemistry, Darmstadt University of Technology, Petersenstr. 22, 64287, Darmstadt, Germany.
Abstract:
Human tissue prokallikrein, a zymogen of the kallikrein-kinin system, circulates in plasma bound to neutrophils. Because plasma kininogens contribute to the assembly of kinin-generating components on blood cells, these proteins were assessed for their ability to complex the kallikrein precursor. Using ligand blot and direct binding assays, biotinylated prokallikrein was found to bind only to high-molecular-weight kininogen and not to the low-molecular-weight form. The interaction was specific, reversible, and saturable yielding an estimated dissociation constant K(D)=690 nM and a 1:1 stoichiometry. Specific kininogen binding of tissue prokallikrein also occurred at physiological plasma protein concentrations. These results provide the first evidence for a novel function of high-molecular-weight kininogen as a binding protein for tissue prokallikrein that could serve to localize the kallikrein precursor on the neutrophil surface.
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