title: Effect of disulfide crosslinking on thermal transitions and chaperone-like activity of human small heat shock protein HspB1.
abstract: Temperature-induced conformational changes of reduced and oxidized HspB1 crosslinked by disulfide bond between single Cys137 of neighboring monomers were analyzed by means of different techniques. Heating of reduced HspB1 was accompanied by irreversible changes of Trp fluorescence, whereas oxidized HspB1 underwent completely reversible changes of fluorescence. Increase of the temperature in the range of 20-70 °C was accompanied by self-association of both reduced and oxidized protein. Further increase of the temperature led to formation of heterogeneous mixture of large self-associated complexes of reduced HspB1 and to formation of smaller and less heterogeneous complexes of oxidized HspB1. Heat-induced changes of oligomeric state of reduced HspB1 were only partially reversible, whereas the corresponding changes of oligomeric state of oxidized HspB1 were almost completely reversible. Oxidation resulted in decrease of chaperone-like activity of HspB1. It is concluded that oxidative stress, inducing formation of disulfide bond, can affect stability and conformational mobility of human HspB1.
authors.affiliation: Department of Biochemistry, School of Biology, Moscow State University, Moscow, 119991, Russian Federation.
authors.forename: Anna S
authors.initials: AS
authors.lastname: Chalova
authors.forename: Maria V
authors.initials: MV
authors.lastname: Sudnitsyna
authors.forename: Pavel I
authors.initials: PI
authors.lastname: Semenyuk
authors.forename: Victor N
authors.initials: VN
authors.lastname: Orlov
authors.forename: Nikolai B
authors.initials: NB
authors.lastname: Gusev
mesh_terms: D003545:Cysteine; D004220:Disulfides / Q000737:chemistry*; D005470:Fluorometry; D055551:HSP27 Heat-Shock Proteins / Q000737:chemistry* / Q000378:metabolism; D006360:Heat-Shock Proteins; D006358:Hot Temperature; D006801:Humans; D008027:Light; D018832:Molecular Chaperones; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011487:Protein Conformation; D011489:Protein Denaturation; D055503:Protein Multimerization; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D013329:Structure-Activity Relationship
chemical_list: D004220:Disulfides; D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D003545:Cysteine