Critical Role of Tyrosine 277 in the Ligand-Binding and Transactivating Properties of Retinoic Acid Receptor α

Mailfait, Sandrine; Bélaïche, Denise; Kouach, Mostafa; Dallery, Nathalie; Chavatte, Philippe; Formstecher, Pierre; Sablonnière, Bernard

doi:10.1021/bi005107d

Cited by 3 publications

(3 citation statements)

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“…Mailfait et al probed the role of tyrosine residues in the binding of the retinoic acid receptor α (RARα) to retinoic acid (RA) with tetranitromethane (Mailfait et al, 2000). The data revealed that only Tyr277 of the three Tyr residues in the ligand-binding domain (LBD) of RARα was not modified.…”

Section: Amino Acid-specific Labels and Examples Of Their Applicmentioning

confidence: 99%

Probing protein structure by amino acid‐specific covalent labeling and mass spectrometry

Mendoza

Vachet

2008

Mass Spectrometry Reviews

328

445

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For many years, amino acid-specific covalent labeling has been a valuable tool to study protein structure and protein interactions, especially for systems that are difficult to study by other means. These covalent labeling methods typically map protein structure and interactions by measuring the differential reactivity of amino acid side chains. The reactivity of amino acids in proteins generally depends on the accessibility of the side chain to the reagent, the inherent reactivity of the label and the reactivity of the amino acid side chain. Peptide mass mapping with ESI- or MALDI-MS and peptide sequencing with tandem MS are typically employed to identify modification sites to provide site-specific structural information. In this review, we describe the reagents that are most commonly used in these residue-specific modification reactions, details about the proper use of these covalent labeling reagents, and information about the specific biochemical problems that have been addressed with covalent labeling strategies.

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Section: Amino Acid-specific Labels and Examples Of Their Applicmentioning

confidence: 99%

Probing protein structure by amino acid‐specific covalent labeling and mass spectrometry

Mendoza

Vachet

2008

Mass Spectrometry Reviews

328

445

View full text Add to dashboard Cite

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“…The mutation of a tyrosine residue for phenylalanine is one of the most conservative mutations that can be made in a protein to probe the role of hydrogen bonding in protein stability. Thus, tyrosine residues of several proteins have been mutated to phenylalanine to investigate the importance of hydrogen bonding to protein stability (3,(7)(8)(9)(10), as well as for studying other roles of tyrosine residues, such as their biological function (9,(11)(12)(13)(14)(15)(16)(17), dimerization (18,19), and contribution to spectroscopic signals (20)(21)(22)(23)(24)(25). While the role of tyrosine hydrogen bonding in maintaining the proper conformation for binding various ligands has been studied (14)(15)(16)26), its more general impact on the local dynamics of the native state and its corresponding functional consequence have received less attention.…”

mentioning

confidence: 99%

“…Thus, tyrosine residues of several proteins have been mutated to phenylalanine to investigate the importance of hydrogen bonding to protein stability (3,(7)(8)(9)(10), as well as for studying other roles of tyrosine residues, such as their biological function (9,(11)(12)(13)(14)(15)(16)(17), dimerization (18,19), and contribution to spectroscopic signals (20)(21)(22)(23)(24)(25). While the role of tyrosine hydrogen bonding in maintaining the proper conformation for binding various ligands has been studied (14)(15)(16)26), its more general impact on the local dynamics of the native state and its corresponding functional consequence have received less attention. Of all the properties of tyrosine residues, their contributions to spectroscopic signals have been investigated the least, especially when compared to the data available for tryptophan residues (27)(28)(29)(30)(31)(32)(33)(34)(35).…”

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confidence: 99%

Variable Contributions of Tyrosine Residues to the Structural and Spectroscopic Properties of the Factor for Inversion Stimulation

Boswell¹,

Mathew²,

Beach³

et al. 2004

Biochemistry

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The diverse roles of tyrosine residues in proteins may be attributed to their dual hydrophobic and polar nature, which can result in hydrophobic and ring stacking interactions, as well as hydrogen bonding. The small homodimeric DNA binding protein, factor for inversion stimulation (FIS), contains four tyrosine residues located at positions 38, 51, 69, and 95, each involved in specific intra- or intermolecular interactions. To investigate their contributions to the stability, flexibility, and spectroscopic properties of FIS, each one was independently mutated to phenylalanine. Equilibrium denaturation experiments show that Tyr95 and Tyr51 stabilize FIS by about 2 and 1 kcal/mol, respectively, as a result of their involvement in a hydrogen bond-salt bridge network. In contrast, Tyr38 destabilizes FIS by about 1 kcal/mol due to the placement of a hydroxyl group in a hydrophobic environment. The stability of FIS was not altered when the solvent-exposed Tyr69 was mutated. Limited proteolysis with trypsin and V8 proteases was used to monitor the flexibility of the C-terminus (residues 71-98) and the dimer core (residues 26-70), respectively. The results for Y95F and Y51F FIS revealed a different proteolytic susceptibility of the dimer core compared to the C-terminus, suggesting an increased flexibility of the latter. DNA binding affinity of the various FIS mutants was only modestly affected and correlated inversely with the C-terminal flexibility probed by trypsin proteolysis. Deconvolution of the fluorescence contribution of each mutant revealed that it varies in intensity and direction for each tyrosine in WT FIS, highlighting the role of specific interactions and the local environment in determining the fluorescence of tyrosine residues. The significant changes in stability, flexibility, and signals observed for the Y51F and Y95F mutations are attributed to their coupled participation in the hydrogen bond-salt bridge network. These results highlight the importance of tyrosine hydrogen-bonding and packing interactions for the stability of FIS and demonstrate the varying roles that tyrosine residues can play on the structural and spectroscopic properties of even small proteins.

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Protein Surface Mapping Using Diethylpyrocarbonate with Mass Spectrometric Detection

2008

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The reliability and information content of diethylpyrocarbonate (DEPC) as a covalent probe of protein surface structure has been improved when used appropriately with mass spectrometric detection. Using myoglobin, cytochrome c, and β-2-microglobulin as model protein systems, we demonstrate for the first time that DEPC can modify Ser and Thr residues in addition to His and Tyr residues. This result expands the capability of DEPC as a structural probe because about 25% of the sequence of the average protein can now be covered using this covalent labeling reagent. In addition, we establish a new approach based on mass spectrometry to ensure the structural integrity of proteins during amino acid-specific covalent labeling reactions. This approach involves monitoring the extent of modification as a function of reagent concentration and allows any small-scale or local perturbations caused by the covalent label to be readily identified and avoided. Results indicate that these dose-response plots are much more reliable and generally applicable probes of possible protein structural changes than fluorescence or circular dichroism spectroscopies. These dose-response plots also provide a means of quantitatively comparing the reactivity of each modified residue. Based on comparisons to known X-ray crystal structures, we find that the solvent accessibility of the reactive atom in the side chain and the presence of a nearby charged residue most affect modification rates. Finally, this improved surface mapping method has been used to determine the effect of Cu(II) binding on the structure of β-2-microglobulin. Results confirm that Cu(II) binds His31, but not any of the other three His residues, and changes the solvent accessibility of residues near His31 and near the N-terminus.

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Critical Role of Tyrosine 277 in the Ligand-Binding and Transactivating Properties of Retinoic Acid Receptor α

Cited by 3 publications

References 0 publications

Probing protein structure by amino acid‐specific covalent labeling and mass spectrometry

Probing protein structure by amino acid‐specific covalent labeling and mass spectrometry

Variable Contributions of Tyrosine Residues to the Structural and Spectroscopic Properties of the Factor for Inversion Stimulation

Protein Surface Mapping Using Diethylpyrocarbonate with Mass Spectrometric Detection

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