Probing Structural Differences in Prion Protein Isoforms by Tyrosine Nitration

Lennon, Christopher; Cox, Holly D.; Hennelly, Scott P.; Chelmo, Sam J.; McGuirl, Michele A.

doi:10.1021/bi0617254

Cited by 24 publications

(28 citation statements)

References 78 publications

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“…Recent reports suggested that YYR sequence in PrPs can be susceptible to the redox changes including labeling by nitrating reagents, if the residues were facing the hydrophilic surface of the PrP protein globule. According to Lennon et al 27, two conserved tyrosine residues on hamster PrP (tyrosines 149 and 150) are insensitive to labeling by nitrating reagents, if PrP was in the normal cellular form. However, these tyrosine residues become reactive after the protein has been converted to the β-oligomeric isoform.…”

Section: Resultsmentioning

confidence: 99%

Free tyrosine and tyrosine-rich peptide-dependent superoxide generation catalyzed by a copper-binding, threonine-rich neurotoxic peptide derived from prion protein

Yokawa¹,

Kagenishi²,

Goto³

et al. 2009

Int. J. Biol. Sci.

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Previously, generation of superoxide anion (O2•-) catalyzed by Cu-binding peptides derived from human prion protein (model sequence for helical Cu-binding motif VNITKQHTVTTTT was most active) in the presence of catecholamines and related aromatic monoamines such as phenylethylamine and tyramine, has been reported [Kawano, T., Int J Biol Sci 2007; 3: 57-63]. The peptide sequence (corresponding to helix 2) tested here is known as threonine-rich neurotoxic peptide. In the present article, the redox behaviors of aromatic monoamines, 20 amino acids and prion-derived tyrosine-rich peptide sequences were compared as putative targets of the oxidative reactions mediated with the threonine-rich prion-peptide. For detection of O2•-, an O2•--specific chemiluminescence probe, Cypridina luciferin analog was used. We found that an aromatic amino acid, tyrosine (structurally similar to tyramine) behaves as one of the best substrates for the O2•- generating reaction (conversion from hydrogen peroxide) catalyzed by Cu-bound prion helical peptide. Data suggested that phenolic moiety is required to be an active substrate while the presence of neither carboxyl group nor amino group was necessarily required. In addition to the action of free tyrosine, effect of two tyrosine-rich peptide sequences YYR and DYEDRYYRENMHR found in human prion corresponding to the tyrosine-rich region was tested as putative substrates for the threonine-rich neurotoxic peptide. YYR motif (found twice in the Y-rich region) showed 2- to 3-fold higher activity compared to free tyrosine. Comparison of Y-rich sequence consisted of 13 amino acids and its Y-to-F substitution mutant sequence revealed that the tyrosine-residues on Y-rich peptide derived from prion may contribute to the higher production of O2•-. These data suggest that the tyrosine residues on prion molecules could be additional targets of the prion-mediated reactions through intra- or inter-molecular interactions. Lastly, possible mechanism of O2•- generation and the impacts of such self-redox events on the conformational changes in prion are discussed.

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Section: Resultsmentioning

confidence: 99%

Free tyrosine and tyrosine-rich peptide-dependent superoxide generation catalyzed by a copper-binding, threonine-rich neurotoxic peptide derived from prion protein

Yokawa¹,

Kagenishi²,

Goto³

et al. 2009

Int. J. Biol. Sci.

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“…Our results revealed a significant increase in covalent modifications of Nrf2 and Keap1 by the aldehyde 4-HNE and increased tyrosine nitration in response to CSE in A549 cells. Tyrosine nitration of proteins has been known to reduce/disrupt protein function, leading to its proteasomal degradation (22,29). Reactive 4-HNE, a major end product of lipid peroxidation formed during oxidative stress, has been known to react with cysteine, histidine, serine, and lysine residues (15).…”

Section: Discussionmentioning

confidence: 99%

Resveratrol induces glutathione synthesis by activation of Nrf2 and protects against cigarette smoke-mediated oxidative stress in human lung epithelial cells

Kode¹,

Saravanan²,

Caito³

et al. 2008

American Journal of Physiology-Lung Cellular and Molecular Phys

391

248

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Nuclear erythroid-related factor 2 (Nrf2), a redox-sensitive transcription factor, is involved in transcriptional regulation of many antioxidant genes, including glutamate-cysteine ligase (GCL). Cigarette smoke (CS) is known to cause oxidative stress and deplete glutathione (GSH) levels in alveolar epithelial cells. We hypothesized that resveratrol, a polyphenolic phytoalexin, has antioxidant signaling properties by inducing GSH biosynthesis via the activation of Nrf2 and protects lung epithelial cells against CS-mediated oxidative stress. Treatment of human primary small airway epithelial and human alveolar epithelial (A549) cells with CS extract (CSE) dose dependently decreased GSH levels and GCL activity, effects that were associated with enhanced production of reactive oxygen species. Resveratrol restored CSE-depleted GSH levels by upregulation of GCL via activation of Nrf2 and also quenched CSE-induced release of reactive oxygen species. Interestingly, CSE failed to induce nuclear translocation of Nrf2 in A549 and small airway epithelial cells. On the contrary, Nrf2 was localized in the cytosol of alveolar and airway epithelial cells due to CSE-mediated posttranslational modifications such as aldehyde/carbonyl adduct formation and nitration. On the other hand, resveratrol attenuated CSE-mediated Nrf2 modifications, thereby inducing its nuclear translocation associated with GCL gene transcription, as demonstrated by GCL-promoter reporter and Nrf2 small interfering RNA approaches. Thus resveratrol attenuates CSE-mediated GSH depletion by inducing GSH synthesis and protects epithelial cells by reversing CSE-induced posttranslational modifications of Nrf2. These data may have implications in dietary modulation of antioxidants in treatment of chronic obstructive pulmonary disease.

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“…EDC, a zero-length cross-linker, combined with tryptic digestion and mass spectrometry, was used to demonstrate cross-links in a PrP dimer (57), and a Gly 90 -Gly 90 cross-link was recently found in PrP Sc using bis(sulfosuccinimidyl) suberate (11.4 Å spacer length), thus providing the first distance constraint for PrP Sc (62). The structures of PrP C , PrP␤, and PrP Sc have also been studied using surface chemical modification with nitration (25,26) or acetylation (25), combined with mass spectrometric detection. Based on our proof-of-principle experiments, proteinase K combined with cleavable cross-linkers will be useful for mass spectrometry-based structural proteomics studies of prions and other difficult-to-digest proteins.…”

Section: Table III Interlysine Cbdps Cross-links Of Prion Proteins Afmentioning

confidence: 99%

Use of Proteinase K Nonspecific Digestion for Selective and Comprehensive Identification of Interpeptide Cross-links: Application to Prion Proteins

Petrotchenko

Serpa

Hardie

et al. 2012

Molecular & Cellular Proteomics

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Chemical cross-linking combined with mass spectrometry is a rapidly developing technique for structural proteomics. Cross-linked proteins are usually digested with trypsin to generate cross-linked peptides, which are then analyzed by mass spectrometry. The most informative cross-links, the interpeptide cross-links, are often large in size, because they consist of two peptides that are connected by a cross-linker. In addition, trypsin targets the same residues as amino-reactive cross-linkers, and cleavage will not occur at these cross-linker-modified residues. This produces high molecular weight crosslinked peptides, which complicates their mass spectrometric analysis and identification. In this paper, we examine a nonspecific protease, proteinase K, as an alternative to trypsin for cross-linking studies. Initial tests on a model peptide that was digested by proteinase K resulted in a "family" of related cross-linked peptides, all of which contained the same cross-linking sites, thus providing additional verification of the cross-linking results, as was previously noted for other post-translational modification studies. The procedure was next applied to the native (PrP C ) and oligomeric form of prion protein (PrP␤). Using proteinase K, the affinity-purifiable CID-cleavable and isotopically coded cross-linker cyanurbiotindipropionylsuccinimide and MALDI-MS cross-links were found for all of the possible cross-linking sites. After digestion with proteinase K, we obtained a mass distribution of the crosslinked peptides that is very suitable for MALDI-MS analysis. Using this new method, we were able to detect over 60 interpeptide cross-links in the native PrP Structural proteomics, which combines protein chemistry methods with modern mass spectrometry techniques for protein and peptides, is an emerging technology in structural biology. One of the tools for modern structural proteomics is chemical cross-linking combined with mass spectrometry (1-5). Cross-linking analysis provides the distance between two cross-linked amino acid residues, whereas mass spectrometry provides information on which residues are connected. The basis of this method is a chemical reaction of the crosslinking reagent with functional groups on a protein, leading to the formation of a covalent bond between each end of the reagent and the protein. The distance between two crosslinked sites is determined by the length of the spacer in the cross-linking reagent. Thus, identification of the cross-linked sites on a protein or a protein complex provides spatial information and distance constraints for the two amino acid residues that are connected by the cross-linker.The use of chemical cross-linking combined with mass spectrometry is a rapidly developing technique for structural proteomics (4, 6 -8). In recent years, we and others have been developing an array of cross-linking and modification reagents, software, and methods specifically designed for protein cross-linking experiments combined with MS analysis (9 -18). For mass spectrometric determina...

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Probing Structural Differences in Prion Protein Isoforms by Tyrosine Nitration

Cited by 24 publications

References 78 publications

Free tyrosine and tyrosine-rich peptide-dependent superoxide generation catalyzed by a copper-binding, threonine-rich neurotoxic peptide derived from prion protein

Free tyrosine and tyrosine-rich peptide-dependent superoxide generation catalyzed by a copper-binding, threonine-rich neurotoxic peptide derived from prion protein

Resveratrol induces glutathione synthesis by activation of Nrf2 and protects against cigarette smoke-mediated oxidative stress in human lung epithelial cells

Use of Proteinase K Nonspecific Digestion for Selective and Comprehensive Identification of Interpeptide Cross-links: Application to Prion Proteins

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