title: Dual effect of arginine on aggregation of phosphorylase kinase.
abstract: Arginine is widely used in biotechnology as a folding enhancer and aggregation suppressor. However, its action on the stability of complexly organized oligomeric proteins, on the one hand, and its role in the formation of supramolecular structures, on the other hand, are poorly known. The investigation is concerned with the effects of arginine on protein-protein interactions using phosphorylase kinase (PhK) as an example. PhK, a 1.3MDa (αβγδ)4 hexadecameric complex, is a Ca(2+)-dependent regulatory enzyme that catalyzes phosphorylation and activation of glycogen phosphorylase b. On the basis of light scattering measurements it was shown that arginine induced aggregation of Ca(2+)-free PhK. On the contrary, when studying Ca(2+), Mg(2+)-induced aggregation of PhK at 37°C, the protective effect of arginine was demonstrated. The data on analytical ultracentrifugation are indicative of disruption of PhK hexadecameric structure under the action of arginine. Though HspB6 and HspB5 suppress aggregation of PhK they do not block the disruption effect of arginine with respect to both forms of PhK (Ca(2+)-free and Ca(2+), Mg(2+)-bound conformers). The dual effect of arginine has been interpreted from view-point of dual behaviour of arginine, functioning both like an osmolyte and a protein denaturant.
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, Moscow 119071, Russia. Electronic address: eronina@inbi.ras.ru.
authors.forename: Tatiana B
authors.initials: TB
authors.lastname: Eronina
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, Moscow 119071, Russia.
authors.forename: Natalia A
authors.initials: NA
authors.lastname: Chebotareva
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, Moscow 119071, Russia.
authors.forename: Nikolai N
authors.initials: NN
authors.lastname: Sluchanko
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, Moscow 119071, Russia.
authors.forename: Valeriya V
authors.initials: VV
authors.lastname: Mikhaylova
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, Moscow 119071, Russia.
authors.forename: Valentina F
authors.initials: VF
authors.lastname: Makeeva
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, Moscow 119071, Russia.
authors.forename: Svetlana G
authors.initials: SG
authors.lastname: Roman
authors.affiliation: Kolsov's Institute of Developmental Biology, Russian Academy of Sciences, Vavilova 26, Moscow 119991, Russia.
authors.forename: Sergey Yu
authors.initials: SY
authors.lastname: Kleymenov
authors.affiliation: Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, Moscow 119071, Russia. Electronic address: kurganov@inbi.ras.ru.
authors.forename: Boris I
authors.initials: BI
authors.lastname: Kurganov
mesh_terms: D000818:Animals; D001120:Arginine / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D050886:HSP20 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D057446:Hydrodynamics; D007477:Ions; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008744:Methylamines / Q000737:chemistry; D010764:Phosphorylase Kinase / Q000737:chemistry* / Q000378:metabolism; D020011:Protective Agents / Q000494:pharmacology; D066329:Protein Aggregates / Q000187:drug effects*; D011817:Rabbits; D013696:Temperature; D014461:Ultracentrifugation
chemical_list: D050886:HSP20 Heat-Shock Proteins; C494382:HSPB6 protein, human; D007477:Ions; D008744:Methylamines; D020011:Protective Agents; D066329:Protein Aggregates; D001120:Arginine; D010764:Phosphorylase Kinase; C005855:trimethyloxamine; D008274:Magnesium; D002118:Calcium
keywords: Aggregation; Arginine; Ca(2+), Mg(2+) ions; HspB5; HspB6; Phosphorylase kinase