title: Structure and properties of avian small heat shock protein with molecular weight 25 kDa.
abstract: The primary structure of chicken small heat shock protein (sHsp) with apparent molecular weight 25 kDa was refined and it was shown that this protein has conservative primary structure 74RALSRQLSSG(83) at Ser77 and Ser81, which are potential sites of phosphorylation. Recombinant wild-type chicken Hsp25, its three mutants, 1D (S15D), 2D (S77D+S81D) and 3D (S15D+S77D+S81D), as well as delR mutant with the primary structure 74RALS-ELSSG(82) at potential sites of phosphorylation were expressed and purified. It has been shown that the avian tissues contain three forms of Hsp25 having pI values similar to that of the wild-type protein, 1D and 2D mutants that presumably correspond to nonphosphorylated, mono- and di-phosphorylated forms of Hsp25. Recombinant wild-type protein, its 1D mutant and Hsp25, isolated from chicken gizzard, form stable high molecular weight oligomeric complexes. The delR, 2D and 3D mutants tend to dissociate and exist in the form of a mixture of high and low molecular weight oligomers. Point mutations mimicking phoshorylation decrease chaperone activity of Hsp25 measured by reduction of dithiothreitol induced aggregation of alpha-lactalbumin, but increase the chaperone activity of Hsp25 measured by heat induced aggregation of alcohol dehydrogenase. It is concluded that avian Hsp25 has a more stable quaternary structure than its mammalian counterparts and mutations mimicking phosphorylation differently affect chaperone activity of avian Hsp25, depending on the nature of target protein and the way of denaturing.
authors.affiliation: Department of Biochemistry, School of Biology, Moscow State University, Moscow 119992, Russia.
authors.forename: Olesya O
authors.initials: OO
authors.lastname: Panasenko
authors.forename: Alim
authors.initials: A
authors.lastname: Seit Nebi
authors.forename: Olesya V
authors.initials: OV
authors.lastname: Bukach
authors.forename: Steve B
authors.initials: SB
authors.lastname: Marston
authors.forename: Nikolai B
authors.initials: NB
authors.lastname: Gusev
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D017931:DNA Primers; D005895:Gizzard, Avian / Q000737:chemistry; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry; D010766:Phosphorylation; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D017931:DNA Primers; D006360:Heat-Shock Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins