The Distefano Research Group uses synthetic organic chemistry in conjunction with a wide variety of biochemical, spectroscopic and computational techniques to study interesting problems at the interface of chemistry and biology.

Chemical Biology

  • Use of peptide libraries to study the specificity of prenylating enzymes 
Wang, Y.-C.; Dozier, J. K.; Beese, L. S.; Distefano, M. D. Rapid Analysis of Protein Farnesyltransferase Substrate Specificity Using Peptide Libraries and Isoprenoid Diphosphate Analogues. ACS Chem. Biol. 2014, 9, 1726-1735DOI: 10.1021/cb5002312
  • Development and use of synthetic isoprenoid analogues for proteomic analysis to identify prenylated proteins involved in disease

Palsuledesai, C. C.; Ochocki, J. D.; Markowski, T. W.; Distefano, M. D. A combination of metabolic labeling and 2D-DIGE analysis in response to a farnesyltransferase inhibitor facilitates the discovery of new prenylated proteins. Mol. Biosys. 2014, 10, 1094-1103. DOI: 10.1039/C3MB70593E

  • Photoaffinity labeling of enzymes involved in protein prenylation

Hahne, K.; Vervacke, J. S.; Shrestha, L.; Donelson, J. L.; Gibbs;, R. A.; Distefano, M. D.; Hrycyna, C. A., Evaluation of Substrate and Inhibitor Binding to Yeast and Human Isoprenylcysteine Carboxyl Methyltransferases (Icmts) using Biotinylated Benzophenone-containing Photoaffinity Probes. Biochem. Biophys. Res. Comm. 2012, 423, 98-103. DOI: 10.1016/j.bbrc.2012.05.089

  • Use of caged forms of prenylation inhibitors to stop prenylation in a spatially controlled manner within cells and tissues

Abate-Pella, D.; Zeliadt, N. A.; Ochocki, J. D.; Warmka, J. K.; Dore, T. M.; Blank, D. M.; Wattenberg, E. V.; Distefano, M. D., Photochemical Modulation of Ras-Mediated Signal Transduction using Caged Farnesyltransferase Inhibitors: Activation via One- and Two-Photon Excitation. ChemBioChem 2012, 13, 1009-1016. DOI: 10.1002/cbic.201290024

Synthetic Methodology

  • Development of methods for the synthesis of peptide libraries containing free C-termini.
Wang, Y.-C.; Distefano, M. D., Solid-Phase Synthesis of C-Terminal Peptide Libraries for Studying the Specificity of Enzymatic Protein Prenylation. Chem. Comm. 2012, 1359-7345. DOI: 10.1039/C2CC31713C
  • Synthetic methods for the preparation of peptides containing C-terminal esters

Diaz-Rodriguez, V.; Mullen, D. G.; Ganusova, E.; Becker, J. M.; Distefano, M. D. Synthesis of Peptides Containing C-Terminal Methyl Esters Using Trityl Side-Chain Anchoring: Application to the Synthesis of a-Factor and a-Factor Analogs. Org. Lett. 2012, 14, 5648-5651. DOI: 10.1021/ol302592v

  • New photoremovable protecting groups for thiol protection

Mahmoodi, M.; Kubo, T.; Abate-Pella, D.; Wissinger, J. E.; Distefano, M. D. "Solvent-Free Synthesis of Bhc-Cl and Application to Cysteine Protection in Solid Phase Peptide Synthesis" In Peptides across the Pacific: Proceedings of the 23rd American Peptide Symposium; Lebl, M., Ed.; Prompt Scientific Publishing: San Diego, 2013, p 248-249.

  • Synthesis of diazirine-containing photoaffinity labeling isoprenoid analogues

Vervacke, J. S.; Funk, A. L.; Wang, Y.-C.; Strom, M.; Hrycyna, C. A.; Distefano, M. D. A Diazirine-Containing Photoactivatable Isoprenoid: Synthesis and Application in Studies with Isoprenylcysteine Carboxyl Methyltransferase. J. Org. Chem. 2014, 79, 1971-1978. DOI: 10.1021/jo402600b

  • Synthesis of isoprenoids containing bioorthogonal functionality

Wollack, J. W.; Monson, B. J.; Dozier, J. K.; Dalluge, J. J.; Poss, K.; Hilderbrand, S. A.; Distefano, M. D. Site-Specific Labeling of Proteins and Peptides with Trans-cyclooctene Containing Handles Capable of Tetrazine Ligation. Chem. Biol. Drug Des. 2014, 84, 140-147. DOI: 10.1111/cbdd.12303

Applications of Prenylation for Biotechnology

  • Using prenylation to prepare site-specifically modified proteins for therapeutic applications
Rashidian, M.; Song, J. M.; Pricer, R. E.; Distefano, M. D., Chemoenzymatic Reversible Immobilization and Labeling of Proteins without Prior Purification. J. Am. Chem. Soc. 2012, 134, 8455–8467. DOI: 10.1021/ja403813b
  • Preparation of defined protein-DNA crosslinks for studying their repair

Yeo, J. E.; Wickramaratne, S.; Khatwani, S.; Wang, Y.-C.; Vervacke, J.; Distefano, M. D.; Tretyakova, N. Y. Synthesis of Site-Specific DNA‚ÄìProtein Conjugates and Their Effects on DNA Replication. ACS Chem. Biol. 2014, 9, 1860-1868. DOI: 10.1021/cb5001795

  • Use of combinations of enzymes, proteins and multifunctional substrates for assembly of complex nanostructures

Rashidian, M.; Kumarapperuma, S. C.; Gabrielse, K.; Fegan, A.; Wagner, C. R.; Distefano, M. D. Simultaneous Dual Protein Labeling Using a Triorthogonal Reagent. J. Am. Chem. Soc. 2013, 135, 16388–16396. DOI: 10.1021/ja403813b