Attractive protein-polymer interactions markedly alter the effect of macromolecular crowding on protein association equilibria.
The dependence of the fluorescence of catalase upon the concentration of added superoxide dismutase (SOD) indicates that SOD binds to saturable sites on catalase. The affinity of SOD for these sites varies with temperature, and with the concentration of each of three nominally inert polymeric additives--dextran 70, Ficoll 70, and polyethylene glycol 2000. At room temperature (25.0 degrees C) and higher, the addition of high concentrations of polymer is found to significantly enhance the affinity of SOD for catalase, but with decreasing temperature the enhancing effect of polymer addition diminishes, and at 8.0 degrees C, addition of polymer has little or no effect on the affinity of SOD for catalase. The results presented here provide the first experimental evidence for the existence of competition between a repulsive excluded volume interaction between protein and polymer, which tends to enhance association of dilute protein, and an attractive interaction between protein and polymer, which tends to inhibit protein association. The net effect of high concentrations of polymer upon protein associations depends upon the relative strength of these two types of interactions at the temperature of measurement, and may vary significantly between different proteins and/or polymers.
Jiao
Ming
M
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, China.
Li
Hong-Tao
HT
Chen
Jie
J
Minton
Allen P
AP
Liang
Yi
Y
D000818:Animals; D001665:Binding Sites; D002374:Catalase / Q000378:metabolism*; D002417:Cattle; D003911:Dextrans / Q000378:metabolism; D019277:Entropy; D005362:Ficoll / Q000378:metabolism; D005453:Fluorescence; D007700:Kinetics; D046911:Macromolecular Substances / Q000378:metabolism*; D008958:Models, Molecular; D011092:Polyethylene Glycols / Q000378:metabolism; D011108:Polymers / Q000378:metabolism*; D011485:Protein Binding; D013482:Superoxide Dismutase / Q000378:metabolism*; D013696:Temperature
D003911:Dextrans; D046911:Macromolecular Substances; D011108:Polymers; D005362:Ficoll; D011092:Polyethylene Glycols; D002374:Catalase; D013482:Superoxide Dismutase