Direct detection of nitrotyrosine-containing proteins using an aniline-based oxidative coupling strategy

Sangsuwan, Rapeepat; Obermeyer, Allie C.; Tachachartvanich, Phum; Palaniappan, Krishnan K.; Francis, Matthew B.

doi:10.1039/c6cc04575h

Cited by 6 publications

(6 citation statements)

References 27 publications

(62 reference statements)

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“…Combining the results from each method allowed unequivocal separation of the modified site from other potentially reactive residues, and the site of the +15 Da modification was confirmed as Y115 (Figure ). By comparison, MS analysis of RNase A modified in solution (using the small nitrating agent tetranitromethane (TNM), and subsequent reduction) was shown to give the 4x modified protein as the major product (Supporting Information, Section 10); further confirming that the observed Tyr residue selectivity could be attributed to the catch‐and‐release approach.…”

Section: Methodsmentioning

confidence: 85%

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A Catch‐and‐Release Approach to Selective Modification of Accessible Tyrosine Residues

et al. 2018

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The tyrosine side chain is amphiphilic leading to significant variations in the surface exposure of tyrosine residues in the folded structure of a native sequence protein. This variability can be exploited to give residue-selective functionalization of a protein substrate by using a highly reactive diazonium group tethered to an agarose-based resin. This novel catch-and-release approach to protein modification has been demonstrated for proteins with accessible tyrosine residues, which are compared with a control group of proteins in which there are no accessible tyrosine residues. MS analysis of the modified proteins showed that functionalization was highly selective, but reactivity was further attenuated by the electrostatic environment of any individual residue. Automated screening of PDB structures allows identification of potential candidates for selective modification by comparison with the accessibility of the tyrosine residue in a benchmark peptide (GYG).

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Section: Methodsmentioning

confidence: 85%

“…To demonstrate the utility of the o ‐aminophenol modification, a fluorophore was conjugated to the catch‐and‐release modified RNase A using an oxidative coupling strategy developed by Francis et al . (Figure ).…”

Section: Methodsmentioning

confidence: 99%

A Catch‐and‐Release Approach to Selective Modification of Accessible Tyrosine Residues

et al. 2018

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“…The cpTMV disks containing ncAAs were then subjected to potassium ferricyanide-mediated oxidative coupling conditions as previously reported by our laboratory for p AF- and 3NY-containing proteins (Figure a). − The accessibility of the mutated amino acids in the cpTMV-S65- p AF and -3NY constructs was first examined through small-molecule couplings. For the 3NY mutant, treatment with sodium dithionite was first required to reduce the nitrophenol to an aminophenol, producing cpTMV-S65-3-aminophenol (cpTMV-S65-3AY).…”

Section: Resultsmentioning

confidence: 99%

Protein-Based Model for Energy Transfer between Photosynthetic Light-Harvesting Complexes Is Constructed Using a Direct Protein–Protein Conjugation Strategy

Bischoff

Hamerlynck

et al. 2023

J. Am. Chem. Soc.

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Photosynthetic organisms utilize dynamic and complex networks of pigments bound within light-harvesting complexes to transfer solar energy from antenna complexes to reaction centers. Understanding the principles underlying the efficiency of these energy transfer processes, and how they may be incorporated into artificial light-harvesting systems, is facilitated by the construction of easily tunable model systems. We describe a protein-based model to mimic directional energy transfer between light-harvesting complexes using a circular permutant of the tobacco mosaic virus coat protein (cpTMV), which self-assembles into a 34-monomer hollow disk. Two populations of cpTMV assemblies, one labeled with donor chromophores and another labeled with acceptor chromophores, were coupled using a direct protein−protein bioconjugation method. Using potassium ferricyanide as an oxidant, assemblies containing o-aminotyrosine were activated toward the addition of assemblies containing p-aminophenylalanine. Both of these noncanonical amino acids were introduced into the cpTMV monomers through amber codon suppression. This coupling strategy has the advantages of directly, irreversibly, and site-selectively coupling donor with acceptor protein assemblies and avoids cross-reactivity with native amino acids and undesired donor−donor or acceptor−acceptor combinations. The coupled donor−acceptor model was shown to transfer energy from an antenna disk containing donor chromophores to a downstream disk containing acceptor chromophores. This model ultimately provides a controllable and modifiable platform for understanding photosynthetic interassembly energy transfer and may lead to the design of more efficient functional light-harvesting materials.

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“…In previous studies, the PTN level in the biological samples was observed up to nanomole level and has significant differences among different samples (plasma, urine or tissue) . In addition, the nitration in tyrosine usually occurs in different kinds of proteins.…”

Section: Methodsmentioning

confidence: 96%

An Azo Coupling Strategy for Protein 3‐Nitrotyrosine Derivatization

Liu

Zhou

et al. 2019

Chemistry A European J

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Herein, as trategy for the selective derivatization of 3-nitrotyrosine-containing proteins using the classic azo coupling reactiona st he keys tep is described. This novel approach featured multiple advantages and was successfully appliedt od etect picomole levelso fp rotein tyrosine nitrationi nb iological samples.Protein tyrosine nitration (PTN), that is, conversion of the tyrosine residue into 3-nitrotyrosine (3-NT)b yr eactive nitrogen species( RNS), is ac ommon protein post-translationalm odification, and is aw ell-establishedb iomarker of nitroxidative stress. [1] Elevationo fP TN level has been observed in multiple pathological conditions, and is closely associatedw itha ging [2] and many diseases, including neurodegeneration, cancera nd cardiovascular diseases. [1c, 3] The occurrence of 3-NT in proteome and the importantr ole of PTN have simulated increasing studies to establish specific and sensitivem ethodsf or detection of 3-NT in biological samples. [1a,d, 4] Traditionald etection methods of 3-NT in biological samples are generally divided to two types. One is based on instrumental analysis, such as HPLC, GC, HPLC-MS, tandem mass spectrometry. [5] However, this kind of detection methods requiresc omplicated sample pretreatment processes. The other is based on immunoassays, such as western blot, immunohistochemistry,i mmunoprecipitation and ELISA,b yu sing 3-NT antibodies. [6] However,t he specificity and affinity of anti-3-NT antibodiesi nr ecognizing different 3-NT-containing proteins are varying as most of thesea ntibodies are produced using 3-NT or specific 3-NT containing proteins as an immunogen without considering the surrounding amino acid sequence and the compatibility of the anti-body,r espectively.I nr ecent years, selective chemical derivatizationo f3 -NT for enriching or detecting PTN has gained increasinga ttention. [4, 5c] In general, 3-NT was first transferred to 3-aminotyrosine (3-AT)b yr educing agents, and subsequently different reagents were employed to selectively capture3 -AT. These3 -AT-reactive partners, such as N-hydroxysuccinimide (NHS)e ster [7] and aldehydes, [8] may capture the 3-AT-containing proteins to facilitatet he following instrumentala nalysis. These reagents react with amines promiscuously,a nd it thus requires blockingo ther protein aminog roups prior to capturing 3-AT. Besides, several coupling reactions that selectively react with 3-ATh ave been developed. For example, Schçneicha nd his colleagues used benzylamined erivatives to achieve at urn-on labeling of PTN under oxidative conditions. [9] Francis and coworkersd eveloped at wo-step chemoselective aniline-based oxidative coupling strategy to detect PTN in cell lysates. [10] However,t he oxidative conditions might cause undesired modifications of oxidative-sensitive residues (such as cysteine and methionine). And based on the mechanism they proposed, [9b, 10b, 11] the benzylamined erivatives or aniline might also react with 3-hydroxytyrosine.N ga nd Wong converted 3-ATt o 3-azidotyrosine, followed by c...

show abstract

Direct detection of nitrotyrosine-containing proteins using an aniline-based oxidative coupling strategy

Cited by 6 publications

References 27 publications

A Catch‐and‐Release Approach to Selective Modification of Accessible Tyrosine Residues

A Catch‐and‐Release Approach to Selective Modification of Accessible Tyrosine Residues

Protein-Based Model for Energy Transfer between Photosynthetic Light-Harvesting Complexes Is Constructed Using a Direct Protein–Protein Conjugation Strategy

An Azo Coupling Strategy for Protein 3‐Nitrotyrosine Derivatization

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