Effects of arginine on kinetics of protein aggregation studied by dynamic laser light scattering and tubidimetry techniques.
Prevention of undesirable protein aggregation is an extremely important strategy in protein science, medicine, and biotechnology. Arginine is one of the most widely used low molecular weight solution additives effective in suppressing aggregation, assisting refolding of aggregated proteins, and enhancing the solubility of aggregation-prone unfolded molecules in vitro. However, the mechanism of suppression of protein aggregation by arginine is not well understood. To address the mechanism, two model systems have been investigated: protection of alcohol dehydrogenase (ADH) and insulin from heat- and dithiothreitol-induced aggregation, respectively, in the presence of arginine. Using dynamic light scattering (DLS) technique, we have demonstrated the concentration-dependent suppression of light scattering intensity of both ADH and insulin aggregates upon addition of arginine to the incubation medium, a significant effect being revealed in the physiological concentration range of arginine (1-10 mM). DLS studies showed that arginine shifted the populations of nanoparticles with higher hydrodynamic radii to the lower ones, suggesting that the preventive effect of arginine on the protein aggregation process arises because it suppresses intermolecular interactions among aggregation-prone molecules. The results of turbidity measurements were also shown to be consistent with these findings.
Lyutova
Elena M
EM
AN Bakh Institute of Biochemistry Russian Academy of Sciences, Leninsky pr., 33, 119071 Moscow, Russia.
Kasakov
Alexei S
AS
Gurvits
Bella Ya
BY
D000426:Alcohol Dehydrogenase / Q000737:chemistry*; D001120:Arginine / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology; D007328:Insulin / Q000737:chemistry*; D007700:Kinetics; D007834:Lasers; D009391:Nephelometry and Turbidimetry; D012542:Scattering, Radiation
D007328:Insulin; D001120:Arginine; D000426:Alcohol Dehydrogenase; D004229:Dithiothreitol