title: Phosphorylation of human small heat shock protein HspB8 (Hsp22) by ERK1 protein kinase.
abstract: A number of phosphomimicking mutants (replacement of Ser/Thr residues by Asp) of human small heat shock protein HspB8 were obtained and phosphorylation of the wild type HspB8 and its mutants by ERK1 kinase was analyzed in vitro. Mutation S159D does not affect phosphorylation, whereas mutations S24D and S27D equally moderately inhibited and mutation T87D strongly inhibited phosphorylation of HspB8. The double mutations S24D/T87D and S27D/T87D induced very strong inhibitory effect and the triple mutations S24D/S27D/T87D completely prevented phosphorylation catalyzed by ERK1. Thus, Ser24 and Thr87, found to be phosphorylated in vivo, are among the sites phosphorylated by ERK1 in HspB8 in vitro. Mutations S24D and T87D affect intrinsic tryptophan fluorescence and susceptibility to chymotrypsinolysis of HspB8. Phosphomimicking mutations and phosphorylation promote concentration-dependent association of HspB8 subunits. Mutations S24D and S27D decrease, whereas mutation T87D increases the chaperone-like activity of HspB8. It is concluded that phosphorylation catalyzed by ERK1 might affect the structure and chaperone-like activity of HspB8 and therefore can be important for regulation of interaction of HspB8 with different target proteins.
authors.affiliation: Department of Biochemistry, School of Biology, Moscow State University, Moscow, Russian Federation.
authors.forename: Anton A
authors.initials: AA
authors.lastname: Shemetov
authors.forename: Alim S
authors.initials: AS
authors.lastname: Seit-Nebi
authors.forename: Nikolai B
authors.initials: NB
authors.lastname: Gusev
mesh_terms: D002850:Chromatography, Gel; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays; D006360:Heat-Shock Proteins / Q000737:chemistry*; D006801:Humans; D048052:Mitogen-Activated Protein Kinase 3 / Q000737:chemistry*; D018832:Molecular Chaperones; D010446:Peptide Fragments / Q000737:chemistry; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: C413214:HSPB8 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D017346:Protein Serine-Threonine Kinases; D048052:Mitogen-Activated Protein Kinase 3