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Analysis of the interaction between human plasma fibronectin and gelatin by affinity electrophoresis
Author(s) -
Kashiwagi Shiro,
Nakamura Kazuyuki,
Takeo Kazusuke,
Takasago Teiichi,
Uchimichi Akihiko,
Ito Haruhide
Publication year - 1991
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150120607
Subject(s) - fibronectin , gelatin , chemistry , urea , hydrogen bond , enthalpy , van der waals force , dissociation (chemistry) , bond dissociation energy , affinity electrophoresis , dissociation constant , chromatography , affinity chromatography , molecule , biochemistry , receptor , enzyme , thermodynamics , organic chemistry , physics , extracellular matrix
The interaction between human plasma fibronectin and gelatin was analyzed by affinity electrophoresis, in which the fibronectin was subjected to electrophoresis in a 4% polyacrylamide gel in the presence and absence of gelatin, as an affinity ligand, and the fibronectin band was stained by an immunoblotting method. The apparent dissociation constants ( K d ) of fibronectin for gelatin were calculated from affinity plots based on the original affinity equation at different pHs, urea concentrations, and temperatures. The fibronectin exhibited much lower affinity in the presence of urea. The K d s at 37°C were 1.49 × 10 −7 M , 2.50 × 10 −6 M , and 3.58 × 10 −6 M with 2 M , 3 M , and 4 M urea, respectively. The van't Hoff plots of K d values against absolute temperature ( T ) showed that the value of log K d decreased in proportion to the increase in the value of 1/ T within the range of 15–50°C. The standard enthalpy, the standard free energy change at 37°C, and the entropy change at 37°C for association were calculated to be −124.7 kJ/mol, −33.23 kJ/mol, and −295.1 J/mol/deg, respectively. These results suggest that a hydrophilic interaction, such as hydrogen bond or van der Waals interaction, plays an important role in the binding of plasma fibronectin to gelatin.