How does dextran sulfate prevent heat induced aggregation of protein? The mechanism and its limitation as aggregation inhibitor.
The effect of dextran sulfate on protein aggregation was investigated to provide the clues of its biochemical mechanism. The interaction between dextran sulfate and BSA varied with the pH values of the solution, which led to the different extent of aggregation prevention by dextran sulfate. Light scattering data with thermal scan showed that dextran sulfate suppressed BSA aggregation at pH 5.1 and pH 6.2, while it had no effect at pH 7.5. Isothermal titration calorimetric analysis suggested that the pH dependency of the role of dextran sulfate on BSA aggregation would be related to the difference in the mode of BSA-dextran sulfate complex formation. Isothermal titration calorimetric analysis at pH 6.2 indicated that dextran sulfate did not bind to native BSA at this pH, but interacted with partially unfolded BSA. While stabilizing native form of protein by the complex formation has been suggested as the suitable mechanism of preventing aggregation, our observation of conformational changes by circular dichroism spectroscopy showed that strong electrostatic interaction between dextran sulfate and BSA rather facilitated the denaturation of BSA. Combining the data from isothermal titration calorimetry, circular dichroism, and dynamic light scattering, we found that the complex formation of the intermediate state of denatured BSA with dextran sulfate is a prerequisite to suppress the aggregation by preventing further oligomerization/aggregation process of denatured protein.
Chung
Kwanghun
K
School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea.
Kim
Juhan
J
Cho
Byung-Kwan
BK
Ko
Byoung-Joon
BJ
Hwang
Bum-Yeol
BY
Kim
Byung-Gee
BG
D002151:Calorimetry; D002942:Circular Dichroism; D016264:Dextran Sulfate / Q000494:pharmacology*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D055672:Static Electricity
D012710:Serum Albumin, Bovine; D016264:Dextran Sulfate