Impact of Hydrogen Peroxide on the Activity, Structure, and Conformational Stability of the Oxidized Protein Repair Enzyme Methionine Sulfoxide Reductase A

Le, Hai-Tuong; Chaffotte, Alain; Demey-Thomas, Emmanuelle; Vinh, Joëlle; Friguet, Bertrand; Mary, Jean

doi:10.1016/j.jmb.2009.07.072

Cited by 21 publications

(9 citation statements)

References 33 publications

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“…The other concern is the sensitivity of proteins toward oxidants, which is difficult to generalize and has to be considered case by case. Although some proteins, including cysteine‐rich proteins, are prone to oxidation and can lose their activity upon treatment with H 2 O 2 , other proteins, such the extracellular matrix proteins, can resist oxidants and are regulated by oxidative stress . Therefore, the herein‐presented method is restricted to proteins that are not sensitive to oxidation.…”

Section: Resultsmentioning

confidence: 99%

“…Although some proteins,i ncluding cysteine-rich proteins,a re prone to oxidation andc an lose their activity upon treatmentw ith H 2 O 2 ,o ther proteins, such the extracellular matrix proteins,can resist oxidants and are regulated by oxidatives tress. [43][44][45][46][47] Therefore, the herein-presented methodi s restricted to proteins that are not sensitive to oxidation. To demonstrate that the oxidation of Co 2 + to Co 3 + with 10 mm H 2 O 2 is compatible with the immobilization of activep roteins, we used His6-tagged ProteinA, which binds IgG antibodies and is widely used to immobilize antibodies in an oriented fashion.…”

Section: Resultsmentioning

confidence: 99%

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Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

Wegner

Schenk

Spatz

2016

Chemistry A European J

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We present the cobalt(III)-mediated interaction between polyhistidine (His)-tagged proteins and nitrilotriacetic acid (NTA)-modified surfaces as a general approach for a permanent, oriented, and specific protein immobilization. In this approach, we first form the well-established Co(2+) -mediated interaction between NTA and His-tagged proteins and subsequently oxidize the Co(2+) center in the complex to Co(3+) . Unlike conventionally used Ni(2+) - or Co(2+) -mediated immobilization, the resulting Co(3+) -mediated immobilization is resistant toward strong ligands, such as imidazole and ethylenediaminetetraacetic acid (EDTA), and washing off over time because of the high thermodynamic and kinetic stability of the Co(3+) complex. This immobilization method is compatible with a wide variety of surface coatings, including silane self-assembled monolayers (SAMs) on glass, thiol SAMs on gold surfaces, and supported lipid bilayers. Furthermore, once the cobalt center has been oxidized, it becomes inert toward reducing agents, specific and unspecific interactions, so that it can be used to orthogonally functionalize surfaces with multiple proteins. Overall, the large number of available His-tagged proteins and materials with NTA groups make the Co(3+) -mediated interaction an attractive and widely applicable platform for protein immobilization.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

Wegner

Schenk

Spatz

2016

Chemistry A European J

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“…aeruginosa is likely to encounter toxic levels of H 2 O 2 during interaction with hosts and in the environment. H 2 O 2 can oxidize both free and bound Met (52). Genetic analyses in other bacteria have shown that inactivation of either msrA or msrB renders the mutants less resistant to H 2 O 2 treatment (45, 53).…”

Section: Resultsmentioning

confidence: 99%

Gene Expression and Physiological Role of Pseudomonas aeruginosa Methionine Sulfoxide Reductases during Oxidative Stress

et al. 2013

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e Pseudomonas aeruginosa PAO1 has two differentially expressed methionine sulfoxide reductase genes: msrA (PA5018) and msrB (PA2827). The msrA gene is expressed constitutively at a high level throughout all growth phases, whereas msrB expression is highly induced by oxidative stress, such as sodium hypochlorite (NaOCl) treatment. Inactivation of either msrA or msrB or both genes (msrA msrB mutant) rendered the mutants less resistant than the parental PAO1 strain to oxidants such as NaOCl and H 2 O 2 . Unexpectedly, msr mutants have disparate resistance patterns when exposed to paraquat, a superoxide generator. The msrA mutant had a higher paraquat resistance level than the msrB mutant, which had a lower paraquat resistance level than the PAO1 strain. The expression levels of msrA showed an inverse correlation with the paraquat resistance level, and this atypical paraquat resistance pattern was not observed with msrB. Virulence testing using a Drosophila melanogaster model revealed that the msrA, msrB, and, to a greater extent, msrA msrB double mutants had an attenuated virulence phenotype. The data indicate that msrA and msrB are essential genes for oxidative stress protection and bacterial virulence. The pattern of expression and mutant phenotypes of P. aeruginosa msrA and msrB differ from previously characterized msr genes from other bacteria. Thus, as highly conserved genes, the msrA and msrB have diverse expression patterns and physiological roles that depend on the environmental niche where the bacteria thrive.

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“…3,10,13 This suggests that typical cellular levels of carbonylation are not likely to disrupt protein structure, which is not in contradiction with experimental studies. 16,18 If carbonylation does not induce major structural destabilization of a typical protein and thus lead to aggregation, as widely assumed, 17,47 how does it then promote the formation of cytotoxic aggregates? First, several studies suggest that some proteins are more susceptible to carbonylation than others, 9,48−50 which may result in a situation where these proteins are completely carbonylated, while the majority of other proteins are still intact, in agreement with our results.…”

Section: Discussionmentioning

confidence: 99%

Microscopic Analysis of Protein Oxidative Damage: Effect of Carbonylation on Structure, Dynamics, and Aggregability of Villin Headpiece

Petrov

Žagrović

2011

J. Am. Chem. Soc.

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One of the most important irreversible oxidative modifications of proteins is carbonylation, the process of introducing a carbonyl group in reaction with reactive oxygen species. Notably, carbonylation increases with the age of cells and is associated with the formation of intracellular protein aggregates and the pathogenesis of age-related disorders such as neurodegenerative diseases and cancer. However, it is still largely unclear how carbonylation affects protein structure, dynamics, and aggregability at the atomic level. Here, we use classical molecular dynamics simulations to study structure and dynamics of the carbonylated headpiece domain of villin, a key actin-organizing protein. We perform an exhaustive set of molecular dynamics simulations of a native villin headpiece together with every possible combination of carbonylated versions of its seven lysine, arginine, and proline residues, quantitatively the most important carbonylable amino acids. Surprisingly, our results suggest that high levels of carbonylation, far above those associated with cell death in vivo, may be required to destabilize and unfold protein structure through the disruption of specific stabilizing elements, such as salt bridges or proline kinks, or tampering with the hydrophobic effect. On the other hand, by using thermodynamic integration and molecular hydrophobicity potential approaches, we quantitatively show that carbonylation of hydrophilic lysine and arginine residues is equivalent to introducing hydrophobic, charge-neutral mutations in their place, and, by comparison with experimental results, we demonstrate that this by itself significantly increases the intrinsic aggregation propensity of both structured, native proteins and their unfolded states. Finally, our results provide a foundation for a novel experimental strategy to study the effects of carbonylation on protein structure, dynamics, and aggregability using site-directed mutagenesis.

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Impact of Hydrogen Peroxide on the Activity, Structure, and Conformational Stability of the Oxidized Protein Repair Enzyme Methionine Sulfoxide Reductase A

Cited by 21 publications

References 33 publications

Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

Gene Expression and Physiological Role of Pseudomonas aeruginosa Methionine Sulfoxide Reductases during Oxidative Stress

Microscopic Analysis of Protein Oxidative Damage: Effect of Carbonylation on Structure, Dynamics, and Aggregability of Villin Headpiece

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