Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Apr 3;141(13):5125-5129.
doi: 10.1021/jacs.9b00837. Epub 2019 Mar 22.

Ruthenium-Catalyzed Redox Isomerizations inside Living Cells

Cristian Vidal  1 , María Tomás-Gamasa  1 , Alejandro Gutiérrez-González  1 , José L Mascareñas  1
Affiliations

Ruthenium-Catalyzed Redox Isomerizations inside Living Cells

Cristian Vidal et al. J Am Chem Soc. .

Abstract

Tailored ruthenium(IV) complexes can catalyze the isomerization of allylic alcohols into saturated carbonyl derivatives under physiologically relevant conditions, and even inside living mammalian cells. The reaction, which involves ruthenium-hydride intermediates, is bioorthogonal and biocompatible, and can be used for the "in cellulo" generation of fluorescent and bioactive probes. Overall, our research reveals a novel metal-based tool for cellular intervention, and comes to further demonstrate the compatibility of organometallic mechanisms with the complex environment of cells.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
(a) Transfer hydrogenations have been developed to either alter the redox status of cells or to reduce abiotic substrates; (b) this work, and ruthenium-hydride intermediates that are likely involved in the reaction.
Figure 2
Figure 2
Reactivity of ruthenium complex in living cells. (a) Ruthenium catalyzed isomerization of allylic alcohol 1g; (b) Fluorescence micrographies of HeLa cells (confocal): (A) cells incubated with substrate 1g (brightfield and green channel); (B) cells incubated with [Ru], washed and treated with substrate 1g (brightfield and green channel); (C) cells incubated with product 2g (brightfield and green channel); (c) CTFC measurements in HeLa, Vero and A549 cells. Reaction conditions: cells were incubated with [Ru] (10 μM) for 30 min, followed by two washings with DMEM and treatment with substrate 1g (100 μM) for 1 h. Error bars represent the standard error of three independent experiments. λex = 385 nm, λem = 520–700 nm. Scale bar: 12.5 μm.
Figure 3
Figure 3
(a) Extracted ion chromatogram of the product 2g generated intracellularly (methanolic extract). HeLa cells pretreated with 10 μM of [Ru] for 50 min were washed twice and incubated with 100 μM of substrate 1g for 30 min/6 h; (b) turnover numbers of intracellular catalysis at 10 and 25 μM of catalyst loading (the quantification was performed considering the total crude material after methanolic extract and two washing steps (DMEM and PBS)); (inset) ICP-MS values of the intracellular accumulation of ruthenium after incubation of cells in DMEM with 10 or 25 μM (in DMSO) for 50 min, double washing with PBS and digestion with HNO3; (c) cytotoxicity studies in HeLa cells. Reaction conditions: cells were incubated with either substrate 1g (100 μM) or product 2g (100 μM) for 6/24 h. Alternatively, cells were mixed with [Ru] (50 μM) for 30 min, washed twice with DMEM and treated with substrate 1g (100 μM) for 6/24 h (labeled as intracellular reaction). Right bar: cells were incubated with [Ru] (50 μM) for 30 min, followed by two washings with DMEM and the toxicity checked after 24 h. Error bars represent the standard error of three independent experiments.
Figure 4
Figure 4
Generation of α,β-unsaturated ketones. (a) Scope of the ruthenium-catalyzed isomerization. aPerformed using 3 (0.2 mmol), solvent (1.0 mL) and [Ru] (2 mol %). b[Ru] (5 mol %), cells lysates 7 mg/mL. (b) Selected biological studies of GSH consumption using the transformation 3b→4b as a model. Reaction conditions: for intracellular catalysis, cells were incubated with [Ru] (50 μM) for 30 min, followed by two washings with DMEM and treatment with substrate 3b (100 μM), for 6 h. For control experiments, cells were incubated with substrate 3b or product 4b (100 μM) for 6 h. Error bars represent the standard error of three independent experiments.
See this image and copyright information in PMC

References

    1. Berg J. M.; Tymoczko J. L.; Stryer L.. Biochemistry; W. H. Freeman: New York, 2002.
    2. Garcia-Viloca M.; Gao J.; Karplus M.; Truhlar D. G. How enzymes work: analysis by modern rate theory and computer simulations. Science 2004, 303, 186–195. 10.1126/science.1088172. - DOI - PubMed
    3. Küchler A.; Yoshimoto M.; Luginbühl S.; Mavelli F.; Walde P. Enzymatic reactions in confined enviornments. Nat. Nanotechnol. 2016, 11, 409–420. 10.1038/nnano.2016.54. - DOI - PubMed
    4. Agarwal P. K. Biophysical perspective on enzyme catalysis. Biochemistry 2019, 58, 438–449. 10.1021/acs.biochem.8b01004. - DOI - PMC - PubMed
    1. Hegedus L. S.Transition metals in the synthesis of complex organic molecules; Wiley-VCH: Weinheim, 1999.
    2. Beller M.; Bolm C.. Transition metals for organic synthesis: building blocks and fine chemicals; Wiley-VCH: Weinheim, 2008.
    3. Hartwig J. F.Organotransition Metal Chemistry: From Bonding to Catalysis; University Science Books: Mill Valley, CA, 2010.
    1. Sasmal P. K.; Streu C. N.; Meggers E. Metal complex catalysis in living biological systems. Chem. Commun. 2013, 49, 1581–1587. 10.1039/C2CC37832A. - DOI - PubMed
    2. Yang M.; Li J.; Chen R. P. Transition metal-mediated bioorthogonal protein chemistry in living cells. Chem. Soc. Rev. 2014, 43, 6511–6526. 10.1039/C4CS00117F. - DOI - PubMed
    3. Vinogradova E. V. Organometallic chemical biology: an organometallic approach to bioconjugation. Pure Appl. Chem. 2017, 89, 1619–1641. 10.1515/pac-2017-0207. - DOI
    4. Rebelein J. G.; Ward T. R. In vivo catalyzed new-to-nature reactions. Curr. Opin. Biotechnol. 2018, 53, 106–114. 10.1016/j.copbio.2017.12.008. - DOI - PubMed
    5. Martínez-Calvo M.; Mascareñas J. L. Organometallic catalysis in biological media and living settings. Coord. Chem. Rev. 2018, 359, 57–79. 10.1016/j.ccr.2018.01.011. - DOI
    6. Ngo A. H.; Bose S.; Do L. H. Intracellular chemistry: integrating molecular inorganic catalysts with living systems. Chem. - Eur. J. 2018, 24, 10584–10594. 10.1002/chem.201800504. - DOI - PubMed
    7. Bai Y.; Chen J.; Zimmerman S. C. Designed transition metal catalysts for intracellular organic synthesis. Chem. Soc. Rev. 2018, 47, 1811–1821. 10.1039/C7CS00447H. - DOI - PubMed
    8. Soldevilla-Barreda J. J.; Metzler-Nolte N. Intracelullar catalysis with selected metal complexes and metallic nanoparticles: advances toward the development of catalytic metallodrugs. Chem. Rev. 2019, 119, 829–869. 10.1021/acs.chemrev.8b00493. - DOI - PubMed
    1. Streu C.; Meggers E. Ruthenium-induced allylcarbamate cleavage in living cells. Angew. Chem., Int. Ed. 2006, 45, 5645–5648. 10.1002/anie.200601752. - DOI - PubMed
    2. Yusop R. M.; Unciti-Broceta A.; Johansson E. M. V.; Sánchez-Martín R. M.; Bradley M. Palladium-mediated intracellular chemistry. Nat. Chem. 2011, 3, 239–243. 10.1038/nchem.981. - DOI - PubMed
    3. Völker T.; Dempwolff F.; Graumann P. L.; Meggers E. Progress towards bioorthogonal catalysis with organometallic compounds. Angew. Chem., Int. Ed. 2014, 53, 10536–10540. 10.1002/anie.201404547. - DOI - PubMed
    4. Sánchez M. I.; Penas C.; Vázquez M. E.; Mascareñas J. L. Metal-catalyzed uncaging of DNA-binding agents in living cells. Chem. Sci. 2014, 5, 1901–1907. 10.1039/C3SC53317D. - DOI - PMC - PubMed
    5. Tonga G. Y.; Jeong Y.; Duncan B.; Mizuhara T.; Mout R.; Das R.; Kim S. T.; Yeh Y.-C.; Yan B.; Hou S.; Rotello V. M. Supramolecular regulation of bioorthogonal catalysis in cells using nanoparticle-embedded transition metal catalysts. Nat. Chem. 2015, 7, 597–603. 10.1038/nchem.2284. - DOI - PMC - PubMed
    6. Tomás-Gamasa M.; Martínez-Calvo M.; Couceiro J. M.; Mascareñas J. L. Transition metal catalysis in the mitochondria of living cells. Nat. Commun. 2016, 7, 12538–12547. 10.1038/ncomms12538. - DOI - PMC - PubMed
    7. Wang J.; Zheng S.; Liu Y.; Zhang Z.; Lin Z.; Li J.; Zhang G.; Wang X.; Li J.; Chen P. R. Palladium-triggered chemical rescue of intracellular proteins via genetically encoded allene-caged tyrosine. J. Am. Chem. Soc. 2016, 138, 15118–15121. 10.1021/jacs.6b08933. - DOI - PubMed
    8. Miller M. A.; Askevold B.; Mikula H.; Kohler R. H.; Pirovich D.; Weissleder R. Nano-palladium is a cellular catalyst for in vivo chemistry. Nat. Commun. 2017, 8, 15906–15918. 10.1038/ncomms15906. - DOI - PMC - PubMed
    9. Stenton B. J.; Oliveira B. L.; Matos M. J.; Sinatra L.; Bernardes G. J. L. A thioether-directed palladium-cleavable linker for targeted bioorthogonal drug decaging. Chem. Sci. 2018, 9, 4185–4189. 10.1039/C8SC00256H. - DOI - PMC - PubMed
    10. Martínez-Calvo M.; Couceiro J. R.; Destito P.; Rodríguez J.; Mosquera J.; Mascareñas J. L. Intracellular deprotection reactions mediated by palladium complexes equipped with designed phosphine ligands. ACS Catal. 2018, 8, 6055–6061. 10.1021/acscatal.8b01606. - DOI - PMC - PubMed
    1. Clavadetscher J.; Hoffmann S.; Lilienkampf A.; Mackay L.; Yusop R. M.; Rider S. A.; Mullins J. J.; Bradley M. Copper catalysis in living systems and in situ drug synthesis. Angew. Chem., Int. Ed. 2016, 55, 15662–15666. 10.1002/anie.201609837. - DOI - PubMed
    2. Li S.; Wang L.; Yu F.; Zhu Z.; Shobaki D.; Chen H.; Wang M.; Wang J.; Qin G.; Erasquin U. J.; Ren L.; Wang Y.; Cai C. Copper-catalyzed click reaction on/in live cells. Chem. Sci. 2017, 8, 2107–2114. 10.1039/C6SC02297A. - DOI - PMC - PubMed
    3. Destito P.; Couceiro J. R.; Faustino H.; López F.; Mascareñas J. L. Ruthenium-catalyzed azide-thioalkyne cycloadditions in aqueous media: a mild, orthogonal, and biocompatible chemical ligation. Angew. Chem., Int. Ed. 2017, 56, 10766–10770. 10.1002/anie.201705006. - DOI - PMC - PubMed
    4. Miguel-Ávila J.; Tomás-Gamasa M.; Olmos A.; Pérez P. J.; Mascareñas J. L. Discrete Cu(I) complexes azide-alkyne annulations of small molecules inside mammalian cells. Chem. Sci. 2018, 9, 1947–1952. 10.1039/C7SC04643J. - DOI - PMC - PubMed

Publication types