title: A protein aggregation based test for screening of the agents affecting thermostability of proteins.
abstract: To search for agents affecting thermal stability of proteins, a test based on the registration of protein aggregation in the regime of heating with a constant rate was used. The initial parts of the dependences of the light scattering intensity (I) on temperature (T) were analyzed using the following empiric equation: I = K(agg)(T-T(0))(2), where K(agg) is the parameter characterizing the initial rate of aggregation and T(0) is a temperature at which the initial increase in the light scattering intensity is registered. The aggregation data are interpreted in the frame of the model assuming the formation of the start aggregates at the initial stages of the aggregation process. Parameter T(0) corresponds to the moment of the origination of the start aggregates. The applicability of the proposed approach was demonstrated on the examples of thermal aggregation of glycogen phosphorylase b from rabbit skeletal muscles and bovine liver glutamate dehydrogenase studied in the presence of agents of different chemical nature. The elaborated approach to the study of protein aggregation may be used for rapid identification of small molecules that interact with protein targets.
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia. eronina@inbi.ras.ru
authors.forename: Tatyana
authors.initials: T
authors.lastname: Eronina
authors.forename: Vera
authors.initials: V
authors.lastname: Borzova
authors.forename: Olga
authors.initials: O
authors.lastname: Maloletkina
authors.forename: Sergey
authors.initials: S
authors.lastname: Kleymenov
authors.forename: Regina
authors.initials: R
authors.lastname: Asryants
authors.forename: Kira
authors.initials: K
authors.lastname: Markossian
authors.forename: Boris
authors.initials: B
authors.lastname: Kurganov
mesh_terms: D000244:Adenosine Diphosphate; D000818:Animals; D001671:Biochemistry / Q000379:methods*; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D003402:Creatine Kinase / Q000737:chemistry; D005969:Glutamate Dehydrogenase / Q000737:chemistry; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000737:chemistry; D007700:Kinetics; D008024:Ligands; D008027:Light; D009391:Nephelometry and Turbidimetry; D010763:Phosphorylase b / Q000737:chemistry; D011489:Protein Denaturation; D055550:Protein Stability; D020836:Protein Structure, Quaternary; D011817:Rabbits; D012542:Scattering, Radiation; D013696:Temperature
chemical_list: D008024:Ligands; D000244:Adenosine Diphosphate; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D005969:Glutamate Dehydrogenase; D010763:Phosphorylase b; D003402:Creatine Kinase