Original Article

Selective catalytic hydrogenation of the N-acyl and uridyl double bonds in the tunicamycin family of protein N-glycosylation inhibitors

Received:
Revised:
Accepted:
Published online:

Abstract

Tunicamycin is a Streptomyces-derived inhibitor of eukaryotic protein N-glycosylation and bacterial cell wall biosynthesis, and is a potent and general toxin by these biological mechanisms. The antibacterial activity is dependent in part upon a π-π stacking interaction between the tunicamycin uridyl group and a specific Phe residue within MraY, a tunicamycin-binding protein in bacteria. We have previously shown that reducing the tunicamycin uridyl group to 5,6-dihydrouridyl (DHU) significantly lowers its eukaryotic toxicity, potentially by disrupting the π-stacking with the active site Phe. The present report compares the catalytic hydrogenation of tunicamycin and uridine with various precious metal catalysts, and describe optimum conditions for the selective production of N-acyl reduced tunicamycin or for tunicamycins reduced in both the N-acyl and uridyl double bonds. At room temperature, Pd-based catalysts are selective for the N-acyl reduction, whereas Rh-based catalysts favor the double reduction to provide access to fully reduced tunicamycin. The reduced DHU is highly base-sensitive, leading to amide ring opening under mild alkaline conditions.

  • Subscribe to The Journal of Antibiotics for full access:

    $305

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    Tunicamycin, (Japan Scientific Societies Press, Tokyo, Japan, (1982).

  2. 2.

    , & Mechanism of action of tunicamycin on theUDP-GlcNAc: dolichyl phosphate GlcNAc-1-phosphate transferase. Biochemistry 18, 2186–2192 (1979).

  3. 3.

    & Modeling bacterial UDP-HexNAc: polyprenol-P HexNAc-1-P transferases. Glycobiology 15, 29–42 (2005).

  4. 4.

    A family of UDP-GlcNAc/MurNAc: polyisoprenol-P GlcNAc/MurNAc-1-Ptransferases. Glycobiology 4, 768–771 (1994).

  5. 5.

    Tunicamycins, streptovirudins, and corynetoxins, a special subclass of Q8 nucleoside antibiotics. J. Nat. Prod. 46, 544–550 (1983).

  6. 6.

    et al. Genome sequences of three tunicamycin-producing streptomyces strains, S. chartreusis NRRL 12338, S. chartreusis NRRL 3882, and S. lysosuperificus ATCC 31396. J. Bacteriol. 193, 7021–7022 (2011).

  7. 7.

    , , & The chemical structures of streptovirudins. J. Antibiot 34, 1631–1632 (1981).

  8. 8.

    , , , & Glycolipid toxins from parasitised annual ryegrass: a comparison with tunicamycin. Biochem. Biophys. Res. Commun. 105, 835–840 (1982).

  9. 9.

    et al. Corynetoxins, causative agents of annual ryegrass toxicity; their identification as tunicamycin group antibiotics. J. Chem. Soc. Commun. 1982, 222–224 (1982).

  10. 10.

    et al. Quinovosamycins: new tunicamycin-type antibiotics in which the α, β-1″,11'-linked N-acetylglucosamine residue is replaced by N-acetylquinovosamine. J. Antibiot. 69, 637–646 (2016).

  11. 11.

    & Biosynthesis of the tunicamycins: a review. J. Antibiot 60, 485–491 (2007).

  12. 12.

    , & Biosynthesis of tunicamycin andmetabolic origin of the 11-carbon dialdose sugar, tunicamine. J. Biol. Chem. 277, 35289–35296 (2002).

  13. 13.

    , , & Dissecting tunicamycin biosynthesis by genome mining: cloning and heterologous expression of a minimal gene cluster. Chem. Sci 1, 581–589 (2010).

  14. 14.

    et al. Characterization of the tunicamycin gene cluster unveiling unique steps involved in its biosynthesis. Protein Cell 1, 1093–1105 (2010).

  15. 15.

    et al. MraY-antibiotic complex reveals details of tunicamycin mode of action. Nat. Chem. Biol. 13, 265–267 (2017).

  16. 16.

    et al. Crystal structure of MraY, an essential membrane enzyme for bacterial cell wall synthesis. Science 341, 1012–1016 (2013). doi:10.1038/ja.2017.101. [Epub ahead of print].

  17. 17.

    et al. Structural insights into inhibition of lipid I production in bacterial cell wall synthesis. Nature 533, 557–560 (2016).

  18. 18.

    et al. Modified tunicamycins with reduced eukaryotic toxicity that enhance the antibacterial activity of β-lactams. J. Antibiot (e-pub ahead of print 27 September 2017; doi:10.1038/ja.2017.101.

  19. 19.

    & Hydrolysis of dihydrouridine and related compounds. Biochemistry 35, 315–320 (1996).

  20. 20.

    & On the specificity of the reduction of transfer ribonucleic acids with sodium borohydride. Eur. J. Biochem. 10, 549–556 (1969).

  21. 21.

    & Selective reduction of yeast transfer ribonucleic acid with sodium borohydride. J. Mol. Biol. 26, 55–66 (1967).

  22. 22.

    , , & The synthesis of dihydrouridine diphosphate glucose. Russ. Chem. Bull. 14, 884–885 (1965).

  23. 23.

    , & The selective photoreduction of uridine in polynucleotides. J. Am. Chem. Soc. 87, 2505–2507 (1965).

  24. 24.

    , , & Photoreduction of uridine and reduction of dihydrouridine with sodium borohydride. J. Am. Chem. Soc. 90, 771–775 (1968).

  25. 25.

    , & Photo-enhanced reduction of carbonyl compounds by sodium borohydride. Tetrahedron Lett. 27, 1157–1160 (1986).

  26. 26.

    & A novel and selective photoisomerization of allylic benzoates. Org. Lett. 3, 3547–3548 (2001).

  27. 27.

    , & Use of specific chemical reagents for detection of modified nucleotides in RNA. J. Nucleic Acids 2011, 1–17 (2011).

  28. 28.

    & Recent advances in antimicrobial nucleoside antibiotics targeting cell wall biosynthesis. Nat. Prod. Rep. 20, 252–273 (2003).

  29. 29.

    et al. Natural and engineered biosynthesis of nucleoside antibiotics in Actinomycetes. J. Ind. Microbiol. Biotechnol. 43, 401–417 (2016).

  30. 30.

    et al. Identification of a napsamycin biosynthesis gene cluster by genome mining. Chembiochem 12, 477–487 (2011).

  31. 31.

    et al. A novel antifungal antibiotic, FR-900848. I. Production, isolation, physico-chemical and biological properties. J. Antibiot. 43, 748–754 (1990).

  32. 32.

    et al. Farnesides A and B, sesquiterpenoid nucleoside ethers from a marine-derived Streptomyces sp., strain CNT-372 from Fiji. J. Nat. Prod. 76, 1815–1818 (2013).

  33. 33.

    et al. Anti-influenza virus compound from Streptomyces sp. RI18. Org. Lett. 12, 4664–4666 (2010).

  34. 34.

    et al. Biosynthesis of the structurally unique polycyclopropanated polyketide-nucleoside hybrid jawsamycin (FR-900848). Angew. Chem. Int. Ed. Engl. 53, 5423–5426 (2014).

Download references

Acknowledgements

We thank Trina Hartman for technical assistance, and Dr Joseph O Rich for pre-review of the manuscript. A provisional patent application (patent no. 62/450,760) has been filed. BY acknowledges the support of the National Natural Science Foundation of China (21372253 and 21432012). Mention of any trade names or commercial products is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. USDA is an equal opportunity provider and employer.

Author information

Affiliations

  1. Agricultural Research Service, US Department of Agriculture, National Center for Agricultural Utilization Research, Peoria, IL, USA

    • Neil PJ Price
    • , Michael A Jackson
    • , Karl E Vermillion
    •  & Judith A Blackburn
  2. State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China

    • Jiakun Li
    •  & Biao Yu

Authors

  1. Search for Neil PJ Price in:

  2. Search for Michael A Jackson in:

  3. Search for Karl E Vermillion in:

  4. Search for Judith A Blackburn in:

  5. Search for Jiakun Li in:

  6. Search for Biao Yu in:

Competing interests

The authors declare no conflict of interest.

Corresponding author

Correspondence to Neil PJ Price.

Supplementary information