Open AccessQuantification of Anti-Aggregation Activity of Chaperones: A Test-System Based on Dithiothreitol-Induced Aggregation of Bovine Serum Albumin
Author(s) -
Vera A. Borzova,
Kira A. Markossian,
Dmitriy A. Kara,
Natalia A. Chebotareva,
Valentina F. Makeeva,
Nikolay B. Poliansky,
Konstantin O. Muranov,
Kurganov Bi
Publication year - 2013
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0074367
Subject(s) - dithiothreitol , bovine serum albumin , chemistry , arginine , protein aggregation , amide , biochemistry , serum albumin , chromatography , biophysics , amino acid , biology , enzyme
The methodology for quantification of the anti-aggregation activity of protein and chemical chaperones has been elaborated. The applicability of this methodology was demonstrated using a test-system based on dithiothreitol-induced aggregation of bovine serum albumin at 45°C as an example. Methods for calculating the initial rate of bovine serum albumin aggregation ( v agg ) have been discussed. The comparison of the dependences of v agg on concentrations of intact and cross-linked α-crystallin allowed us to make a conclusion that a non-linear character of the dependence of v agg on concentration of intact α-crystallin was due to the dynamic mobility of the quaternary structure of α-crystallin and polydispersity of the α-crystallin–target protein complexes. To characterize the anti-aggregation activity of the chemical chaperones (arginine, arginine ethyl ester, arginine amide and proline), the semi-saturation concentration [L] 0.5 was used. Among the chemical chaperones studied, arginine ethyl ester and arginine amide reveal the highest anti-aggregation activity ([L] 0.5 = 53 and 58 mM, respectively).