Binding of adenosine to pendant phenylboronate groups of thermoresponsive copolymer: a quantitative study

Binding of adenosine to the thermosensitive copolymer of N-isopropylacrylamide and 3-(acrylamido)aminophenylboronic acid (82:18, m ($) over bar (n)=47000 g center dot mol(-1)) was studied by equilibrium dialysis at 22 degrees C and 37 degrees C, in a 0.1 m glycine buffer containing 0.1 m NaCl at pH 9.2. The copolymer exhibited a the phase transition temperature (T-p) of 26.5 degrees C under the above conditions. At 22 degrees C the binding of adenosine to the water-soluble copolymer was well described by a Langmuir model, accounting for preferential ionisation of the boronate-nucleoside complexes and, therefore, restricted reactivity of the rest of boronates. At saturation, the copolymer contained 38% of its phenylboronic acid groups in the form of complexes, whereas the association constant was 1400 m(-1). At 37 degrees C no binding of adenosine to thermally precipitated copolymer was found, presumably owing to interaction of the phenylboronates with hydrophobic segments of polyNIPAM. At high loading of the copolymer by the reversibly bound adenosine the T (p) steeply increases with increasing fraction of the phenylboronate-adenosine complexes in the chains. The increase of the T-p observed above the saturating adenosine concentration (> 1 x 10(-3) m, 22 degrees C) very probably testifies to competition of the nucleoside with hydrophobic polyNIPAM segments for binding to the pendant phenylboronates