The mechanism of thermal aggregation of glutamate dehydrogenase. The effect of chemical chaperones.

Autor: Borzova VA; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia. Electronic address: vera.a.borzova@gmail.com., Chebotareva NA; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia., Sluchanko NN; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia., Kleymenov SY; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia; Koltzov Institute of Developmental Biology of the Russian Academy of Sciences, Vavilova 26, Moscow, 119991, Russia., Markossian KA; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia., Kurganov BI; Bach Institute of Biochemistry, Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, Leninsky pr. 33, Moscow, 119071, Russia.
Jazyk: angličtina
Zdroj: Biochimie [Biochimie] 2022 Apr; Vol. 195, pp. 27-38. Date of Electronic Publication: 2022 Jan 15.
DOI: 10.1016/j.biochi.2022.01.004
Abstrakt: Chemical chaperones are low-molecular compounds counteracting protein aggregation. Understanding of the mechanism of their effects is key to their potential use in biotechnology. The aggregation of bovine liver glutamate dehydrogenase (GDH) was studied at 40 °C and 50 °C using dynamic light scattering, analytical ultracentrifugation, size-exclusion chromatography and differential scanning calorimetry. At 40 °C the GDH aggregation proceeds through the slow stages of hexamer dissociation and formation of small oligomeric aggregates. At 50 °C these stages are transient. The rate-limiting stage of the overall aggregation process is unfolding of the protein molecule; the order of aggregation with respect to protein, n = 1. The test system based on GDH aggregation at 50 °C was used to quantify the anti-aggregation activity of chemical chaperones by comparing their half-saturation concentrations [L] 0.5 . Arginine ethyl ester had the highest anti-aggregation activity, with [L] 0.5  = 4 ± 1 mM. For other additives, [L] 0.5 was 22 ± 1 mM (arginine), 18 ± 1 mM (argininamide) and 95 ± 12 mM (proline). Arginine at concentrations up to 300 mM, argininamide at concentrations higher than 300 mM and arginine ethyl ester at concentrations higher than 500 mM enhance aggregate-aggregate sticking. These results explain the mechanism of heat-induced GDH aggregation and its peculiarities at different temperatures or in the presence of chemical chaperones.
Competing Interests: Declaration of competing interest The authors declare that they have no competing interests.
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Databáze: MEDLINE
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