1-Methyl-4-aryl-urazole (MAUra) labels tyrosine in proximity to ruthenium photocatalysts

Sato, Shinichi; Hatano, Kensuke; Tsushima, Michihiko; Nakamura, Hiroyuki

doi:10.1039/c8cc02891e

Cited by 67 publications

(83 citation statements)

References 41 publications

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“…We were able to detect PTAD labels for all previously published sites in BSA except for Y393 21 . We also found previously unreported PTAD labeling at residues Y173, Y179, Y180, and Y520 19,[21][22][23] . We interpreted this to be a result of improved sensitivity by LC-MS/MS.…”

Section: Ptad Labeling Is Distinct For Alternate Folded States In a Psupporting

confidence: 73%

Discriminating changes in protein structure using PTAD conjugation to tyrosine

Moinpour

Barker

Guzman

et al. 2020

Preprint

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Many proteins of high interest to biology and biomedical research have few options that can provide even simple insights into structure, such as whether the protein conformation has changed between two states. Tyrosine is an abundant amino acid that balances hydrophobic and hydrophilic properties such that it can be found at protein surfaces, binding interfaces, and buried in a protein core. The reactive compound 4-phenyl-3H-1,2,4-triazole-3,5(4H)-dione (PTAD) can conjugate the phenolic side chain of tyrosine. We hypothesized that individual protein conformations could be distinguished by changes induced to tyrosine reactivity with PTAD. We quantified for a wellfolded protein, bovine serum albumen (BSA), the complex distribution of PTAD labeling using LC-MS/MS. As a model conformational switch in BSA, we chose to label unfolded BSA using urea. We found that changes in PTAD reactivity for many tyrosine residues was easily quantified and several sites were highly sensitive to the changes produced by unfolding the protein. This study suggests that the reaction of PTAD to tyrosine may allow for the measurement of a covalent fingerprint that can discriminate conformational states in a protein.

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Section: Ptad Labeling Is Distinct For Alternate Folded States In a Psupporting

confidence: 73%

Discriminating changes in protein structure using PTAD conjugation to tyrosine

Moinpour

Barker

Guzman

et al. 2020

Preprint

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“…Based on chemistry established by Kodadek et al [30][31][32] , the group of Nakamura developed a photocatalytic LDL method based on local single-electron transfer (SET) mediated by a Ru(II)(bpy) 3 photocatalyst tethered to a small molecule ( Fig. 1a) [33][34][35][36][37] . Irradiation of the ruthenium complex with visible light results in an excited state [Ru(II) (bpy) 3 ]* complex of relatively long lifetime (~1 ms) that can function as electron donor or electron acceptor 38 .…”

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confidence: 99%

Photocatalytic proximity labelling of MCL-1 by a BH3 ligand

et al. 2019

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Ligand-directed protein labelling allows the introduction of diverse chemical functionalities onto proteins without the need for genetically encoded tags. Here we report a method for the rapid labelling of a protein using a ruthenium-bipyridyl (Ru(II)(bpy) 3 )-modified peptide designed to mimic an interacting BH3 ligand within a BCL-2 family protein-protein interactions. Using sub-stoichiometric quantities of (Ru(II)(bpy) 3 )-modified NOXA-B and irradiation with visible light for 1 min, the anti-apoptotic protein MCL-1 can be photolabelled with a variety of functional tags. In contrast with previous reports on Ru(II)(bpy) 3 -mediated photolabelling, tandem mass spectrometry experiments reveal that the labelling site is a cysteine residue of MCL-1. MCL-1 can be labelled selectively in mixtures with other proteins, including the structurally related BCL-2 member, BCL-x L . These results demonstrate that proximityinduced photolabelling is applicable to interfaces that mediate protein-protein interactions, and pave the way towards future use of ligand-directed proximity labelling for dynamic analysis of the interactome of BCL-2 family proteins.

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“…[19] However,i ti sn ot clear at this stage whether the reaction is initiated with the oxidation of the phenyldiamine, which then reacts with tyrosine, or vice versa. [25] The methodology was showcased with the selective labeling of carbonic anhydrase in crude cell lysates using as ulfonamide ligand that binds specifically to this protein. The reactionw as performed with1mm concentrationo f the ligand-[Ru(bpy) 3 ] 2 + conjugate and 500 mm of the tyrosyl trapping reagents to yield as elective labeling of carbonic anhydrase.Af urther refinement of this methodology was recently reported with the use of 1-methyl-4-arylazole derivatives.…”

Section: Oxidative Receptor Labeling Reactionsmentioning

confidence: 99%