Redox switch of hsp33 has a novel zinc-binding motif.
The chaperone activity of the heat shock protein Hsp33 is regulated by reversible disulfide bond formation. Oxidized Hsp33 is active, and reduced Hsp33 is inactive. We show that zinc binding is essential for the function of this redox switch. Our results reveal that Hps33 contains a new, high affinity (K(a) > 10(17) m(-)(1)), zinc-binding motif in the form Cys-X-Cys-X(27-32)-Cys-X-X-Cys. All four conserved cysteines within this motif act to coordinate a single zinc atom. Experiments where reduced wild type Hsp33 is reconstituted with cobalt or cadmium demonstrate that the metal-coordinating cysteines are present as highly reactive thiolate anions. This ionization may allow for the fast and successful activation of the chaperone function of Hsp33 upon incubation in oxidizing agents.
Jakob
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U
Department of Biology, University of Michigan, Ann Arbor, Michigan 48109-1048, USA. ujakob@biology.lsa.umich.edu
Eser
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Bardwell
J C
JC
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001426:Bacterial Proteins; D001665:Binding Sites; D017124:Conserved Sequence; D003545:Cysteine; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc / Q000378:metabolism*; D016335:Zinc Fingers
D001426:Bacterial Proteins; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D015032:Zinc; D003545:Cysteine