The chaperone-like activity of a small heat shock protein is lost after sulfoxidation of conserved methionines in a surface-exposed amphipathic α-helix

Härndahl, Ulrika; Kokke, Bas P.A.; Gustavsson, Niklas; Linse, Sara; Berggren, Kristina; Tjerneld, Folke; Boelens, Wilbert C.; Sundby, Cecilia

doi:10.1016/s0167-4838(00)00280-6

Cited by 41 publications

(37 citation statements)

References 39 publications

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“…For example, Hsp21, another stress-induced chaperone, converts 6 of its 8 Met residues to sulfoxides when subjected to high concentrations of H 2 O 2 (7 mM for 2 h at 37°C) (57). Even treatment with 1.5 mM H 2 O 2 was sufficient to abolish activity (57,58). In contrast, GroEL treated with 10 mM H 2 O 2 retained significant activity (60%) in our study and retained full activity in studies by Wang et al (59).…”

Section: Discussionmentioning

confidence: 99%

Potential Role of Methionine Sulfoxide in the Inactivation of the Chaperone GroEL by Hypochlorous Acid (HOCl) and Peroxynitrite (ONOO–)

Khor

Fisher

Schöneich

2004

Journal of Biological Chemistry

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GroEL is an Escherichia coli molecular chaperone that functions in vivo to fold newly synthesized polypeptides as well as to bind and refold denatured proteins during stress. This protein is a suitable model for its eukaryotic homolog, heat shock protein 60 (Hsp60), due to the high number of conserved amino acid sequences and similar function. Here, we will provide evidence that GroEL is rather insensitive to oxidants produced endogenously during metabolism, such as nitric oxide ( ⅐ NO) or hydrogen peroxide (H 2 O 2 ), but is efficiently modified and inactivated by reactive species generated by phagocytes, such as peroxynitrite (ONOO ؊ ) and hypochlorous acid (HOCl). For the exposure of 17.5 M GroEL to 100 -250 M HOCl, the major pathway of inactivation was through the oxidation of methionine to methionine sulfoxide, established through mass spectrometric detection of methionine sulfoxide and the reactivation of a significant fraction of inactivated GroEL by the enzyme methionine sulfoxide reductase B/A (MsrB/A). In addition to the oxidation of methionine, HOCl caused the conversion of cysteine to cysteic acid and this product may account for the remainder of inactivated GroEL not recoverable through MsrB/A. In contrast, HOCl produced only negligible yields of 3-chlorotyrosine. A remarkable finding was the conversion of Met 111 and Met 114 to Met sulfone, which suggests a rather low reduction potential of these 2 residues in GroEL. The high sensitivity of GroEL toward HOCl and ONOO ؊ suggests that this protein may be a target for bacterial killing by phagocytes.GroEL and its eukaryotic analog, heat shock protein 60 (Hsp60), 1 are highly sequence-related members of the Group I subclass of chaperonin 60 (Cpn60) (1). These proteins assist the folding of newly synthesized polypeptides (GroEL) or translocated preproteins (mitochondrial Hsp60). The functional unit of GroEL (and of most Cpn60 proteins) is a sandwich of two heptameric rings, which are stacked end to end. Depending on the protein substrate, different ligands such as K ϩ , Mg 2ϩ , ATP, and the cofactor GroES (or Hsp10) may be required for proper folding. Following the trapping of an unfolded or misfolded protein substrate in the hydrophobic interior of GroEL, the binding of ATP and GroES causes a conformational transition, which changes the interior surface properties from hydrophobic to hydrophilic, thus triggering protein folding (1, 2). Mammalian Hsp60 differs from GroEL in that it forms stable and functional heptameric rings in the absence of ATP and its cofactor Hsp10 (3). Our rationale for investigating the oxidative inactivation of GroEL is 2-fold: (i) its potential involvement in bacterial killing by phagocytes and (ii) a potential role for its analog, Hsp60, in an inflammatory and proapoptotic response during cardiovascular disease, as described below.Hsp60 proteins play an important role in the cellular protection against oxidative stress (4, 5). Studies with mutant strains of Saccharomyces cerevisiae exposed to various oxidants show that a decre...

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Section: Discussionmentioning

confidence: 99%

Potential Role of Methionine Sulfoxide in the Inactivation of the Chaperone GroEL by Hypochlorous Acid (HOCl) and Peroxynitrite (ONOO–)

Khor

Fisher

Schöneich

2004

Journal of Biological Chemistry

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“…This part of Hsp21 is a disordered, methionine-rich domain and has previously been suggested to be substratebinding (Harndahl et al 2001). For other sHsps, the Nterminal region has also previously been shown to be implicated in substrate-binding (Jaya et al 2009;Sharma et al 2000).…”

Section: Frequency Of the Different Hsp21 And Mdh Residues Involved Imentioning

confidence: 98%

Probing the transient interaction between the small heat-shock protein Hsp21 and a model substrate protein using crosslinking mass spectrometry

Lambert

Rutsdottir

Hussein

et al. 2013

Cell Stress and Chaperones

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Small heat-shock protein chaperones are important players in the protein quality control system of the cell, because they can immediately respond to partially unfolded proteins, thereby protecting the cell from harmful aggregates. The small heat-shock proteins can form large polydisperse oligomers that are exceptionally dynamic, which is implicated in their function of protecting substrate proteins from aggregation. Yet the mechanism of substrate recognition remains poorly understood, and little is known about what parts of the small heat-shock proteins interact with substrates and what parts of a partially unfolded substrate protein interact with the small heat-shock proteins. The transient nature of the interactions that prevent substrate aggregation rationalize probing this interaction by crosslinking mass spectrometry. Here, we used a workflow with lysine-specific crosslinking and offline nano-liquid chromatography matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry to explore the interaction between the plant small heat-shock protein Hsp21 and a thermosensitive model substrate protein, malate dehydrogenase. The identified crosslinks point at an interaction between the disordered N-terminal region of Hsp21 and the C-terminal presumably unfolding part of the substrate protein.

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“…Evidence is accumulating to suggest an important role for heat shock proteins/molecular chaperones in stress resistance in plant and animal systems (Gustavsson et al 1999;Wehmeyer and Vierling 2000;Harndahl et al 2001). Increased expression of HSPs of the 70, 101 and sHSP classes were observed in drought acclimated rooted cut- Mullineaux et al 2000).…”

Section: Molecular Chaperones Interact To Protect Against Heat and Wamentioning

confidence: 99%

“…These HSPs may prevent loss of conformation in low-water conditions and may be important in peroxisomes. Denatured substrate proteins are bound to the sHSP oligomers in vitro presumably by hydrophobic regions (Lee and Vierling 1998;Harndahl et al 2001). It is thought that the sHSPs act to bind denatured proteins and to maintain them in a state that allows for ATP-dependent refolding by larger HSPs/molecular chaperones (Lee and Vierling 2000).…”

Section: Molecular Chaperones Interact To Protect Against Heat and Wamentioning

confidence: 99%

Oxidative Stress and Acclimation Mechanisms in Plants

Grene

2002

The Arabidopsis Book

116

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The chaperone-like activity of a small heat shock protein is lost after sulfoxidation of conserved methionines in a surface-exposed amphipathic α-helix

Cited by 41 publications

References 39 publications

Potential Role of Methionine Sulfoxide in the Inactivation of the Chaperone GroEL by Hypochlorous Acid (HOCl) and Peroxynitrite (ONOO–)

Potential Role of Methionine Sulfoxide in the Inactivation of the Chaperone GroEL by Hypochlorous Acid (HOCl) and Peroxynitrite (ONOO–)

Probing the transient interaction between the small heat-shock protein Hsp21 and a model substrate protein using crosslinking mass spectrometry

Oxidative Stress and Acclimation Mechanisms in Plants

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