Combined action of chemical chaperones on stability, aggregation and oligomeric state of muscle glycogen phosphorylase b

• Published in: International journal of biological macromolecules 2022-04, Vol.203, p.406-416
• Main Authors: Eronina, Tatiana B., Mikhaylova, Valeriya V., Chebotareva, Natalia A., Kleymenov, Sergey Y., Pivovarova, Anastasia V., Kurganov, Boris I.
• Format: Article
• Language: English
• Subjects: Aggregation; Denaturation; Glycogen phosphorylase b; Glycogen Phosphorylase, Muscle Form - chemistry; Mixture of chemical chaperones; Molecular Chaperones; Muscles - metabolism; Phosphorylase b; Protein Aggregates; Ultracentrifugation
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2022.01.106

Chemical chaperones are a class of small molecules, which enhance protein stability, folding, inhibit protein aggregation, and are used for long-term storage of therapeutic proteins. The combined action of chemical chaperones trehalose, betaine and lysine on stability, aggregation and oligomeric state of muscle glycogen phosphorylase b (Phb) has been studied. Dynamic light scattering data indicate that the affinity of trehalose to Phb increased in the presence of betaine or lysine at both stages (stage of nucleation and aggregate growth) of enzyme aggregation at 48 °C, in contrast, the affinity of betaine to the enzyme in the presence of lysine remained practically unchanged. According to differential scanning calorimetry and analytical ultracentrifugation data, the mixture of trehalose and betaine stabilized Phb stronger than either of them in total. Moreover, the destabilizing effect of lysine on the enzyme was almost completely compensated by trehalose and only partially by betaine. The main protective effect of the mixtures of osmolytes and lysine is associated with their influence on the dissociation/denaturation stage, which is the rate-limiting one of Phb aggregation. Thus, a pair of chaperones affects the stability, oligomeric state, and aggregation of Phb differently than individual chaperones. •The affinity of Tre to Phb is increased in 1.4–1.7 times in the presence of Bet or Lys at 48 °C.•The affinity of Bet to Phb in the presence of Lys remains practically unchanged at 48 °C.•Tre and Bet stabilize the Phb dimer enhancing the protective effect of each other.•The negative effect of Lys on Phb can be removed by increasing concentration of Tre or Bet.•A mixture of chaperones has a different effect on Phb than that of individual chaperones.