A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

The effect of the intersubunit disulfide bond on the structural and functional properties of the small heat shock protein Hsp25

Julkaisun tekijät: Zavialov A, Benndorf R, Ehrnsperger M, Zav'yalov V, Dudich I, Buchner J, Gaestel M


Julkaisuvuosi: 1998

Journal: International Journal of Biological Macromolecules


Lehden akronyymi: INT J BIOL MACROMOL

Volyymi: 22

Julkaisunumero: 3-4

Sivujen määrä: 11

ISSN: 0141-8130

DOI: http://dx.doi.org/10.1016/S0141-8130(98)00014-2

The murine small heat shock protein Hsp25 carries a single cysteine residue in position 141 of its amino acid sequence. Interestingly, Hsp25 can exist within the cell as covalently bound dimer which is linked by an intermolecular disulfide bond between two monomers. Oxidative stress caused by treatment of the cells with diamide, arsenite, or hydrogen peroxide leads to an increase in Hsp25-dimerisation which can be blocked by simultaneous treatment with reducing agents. Recombinant Hsp25 was prepared in an oxidized dimeric (oxHsp25) and reduced monomeric (redHsp25) form. The two species were compared with regard to secondary structure, stability, oligomerization properties and their chaperone activity. It is demonstrated by CD measurements in the far W region that there are no significant differences in the secondary structure and temperature- or pH-stability of oxHsp25 and redHsp25. However, according to CD measurements in the near UV region an increase in the asymmetry of the microenvironment of aromatic residues in oxHsp25 is observed. Furthermore, an increase in stability of the hydrophobic environment of the tryptophan residues mainly located in the N-terminal domain of the protein against urea denaturation is detected in oxHsp25. Both reduced and oxidized Hsp25 form oligomeric complexes of similar size and stability against detergents and both species prevent thermal aggregation of citrate synthase and assist significantly in oxaloacetic acid-induced refolding of the enzyme. Hence, the overall secondary structure, the degree of oligomerization and the chaperone activity of Hsp25 seem independent of the formation of the intermolecular disulfide bond and only the stability of the hydrophobic N-terminal part of the molecule is influenced by formation of this bound. The obtained data do not exclude the possible involvement of dimerization of this protein in other cellular functions, e.g. in intracellular sulfhydryl-buffering or in the protection of actin filaments from fragmentation upon oxidative stress. (C) 1998 Elsevier Science B.V. All rights reserved.

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