Rudi Fasan

Rudi Fasan

Associate Professor of Chemistry

Ph.D. 2005, University of Zurich, Switzerland

Bioorganic Chemistry and Chemical Biology

The research interests of our group lie in the areas of bioorganic chemistry, chemical biology, and biomolecular engineering and evolution. Our laboratory integrates methods and principles of organic chemistry, molecular biology, and molecular evolution to develop novel chemo-biosynthetic and chemo-enzymatic approaches to the discovery of biologically active molecules. Our goal is to investigate and apply these methodologies toward the development of chemical agents useful for probing cell signaling pathways and controlling biomolecular interactions implicated in cancer development.

A major project involves the development of new strategies to direct the synthesis, diversification, and evolution of macrocyclic peptide-based molecules as potent and selective modulators of protein-protein interactions (PPIs). PPIs are implicated in all cellular processes from signal transduction to gene regulation, cell proliferation and apoptosis. Chemical agents capable of targeting PPIs with high potency and selectivity can provide invaluable tools for the study of complex cellular pathways and useful starting points for development of new therapeutics, but the development of such compounds has constituted a major challenge in chemical biology and drug discovery. Our group has pioneered methods to generate Macrocyclic Organo-Peptide Hybrids (MOrPHs) via the modular assembly of synthetic organic moieties with genetically encoded polypeptide precursors. These molecules constitute attractive molecular scaffolds to mediate specific and high-affinity recognition of a target protein as they combine a high degree of chemical complexity with a compact and conformationally rigid architecture. In addition, the MOrPH approach enables to couple the versatility of chemical synthesis with the advantages of genetic encoding and power of genetic mutagenesis, providing unique opportunities for the creation and screening of highly diverse chemical libraries and the molecular evolution of organo-peptide macrocycles with tailored protein-binding selectivity. Our group is exploring the potential of this new class of macrocyclic structures to tackle challenging molecular recognition problems, such as the selective and efficient disruption of protein-protein and protein-DNA complexes of therapeutic relevance and the specific recognition of structural homologues and post-translational isoforms of human proteins. An integral component of these studies is the application of a variety of biophysical techniques (e.g. Surface Plasmon Resonance, NMR and fluorescence spectroscopy) and biological assays for the characterization of the conformational and binding properties and biological activity of these compounds.

Another major area of interest is concerned with the design and development of efficientbiological catalysts for the selective functionalization of aliphatic C—H bonds. The selective functionalization of unactivated C(sp3)—H bonds represents a most valuable but also one of the most challenging transformations in organic chemistry. Our group is investigating the use of engineered cytochrome P450s as well as other heme-containing proteins (e.g. myoglobin) as catalytic platforms for the direct conversion of C(sp3)—H bonds into C—O, C—N, and C—C bonds. With the latter two, we aim at expanding the reactivity scope of these protein-based catalysts beyond that exhibited by naturally occurring enzymes. As part of these projects, we are developing systematic, rationally-driven approaches to predict the reactivity of these P450- and myoglobin-based catalysts via the combination of high-throughput screening methods and computational tools. We are also exploring new protein engineering strategies to enable rapid fine-tuning of the regio- and stereoselectivity of these enzymes. The ultimate goal of these studies is to develop efficient and systematic approaches to expedite the development of C—H functionalization biocatalysts with tailored activity and site-selectivity for synthetic applications. Finally, the scope and synthetic value of these methodologies is being investigated through the synthesis and functional elaboration of complex natural product scaffolds of medical interest via chemoenzymatic synthesis.

All our projects involve the synergistic integration of rational design, chemical synthesis, protein chemistry, and molecular evolution methods toward the development of enabling molecular discovery platforms of practical and broad utility. Members of the Fasan group have thus the opportunity to receive training in these areas and conduct interdisciplinary research at the intersection of chemistry, biology, and biophysics.

Selected Publications

Tyagi V, Sreenilayam G, Bajaj P, Tinoco A, Fasan* R  “Asymmetric synthesis of allylic and allenyl sulfides via a biocatalytic Doyle-Kirmse reaction,”  Angewandte Chemie International Edition  2016in press.
Tyagi V, Alwaseem H, Fasan* R  “Chemoenzymatic synthesis and antileukemic activity of novel C9- and C14-functionalized parthenolide analogs,”  Bioorganic and Medicinal Chemistry  201624(17)3876-86.
Giovani S, Alwaseem H, Fasan* R  “Aldehyde and ketone synthesis via P450-catalyzed oxidative deamination of alkyl azides,”  ChemCatChem  20168(16)2609-2613.
Frost JR, Wu Z, Lam YC, Owens AD, Fasan* R  “Side-chain-to-tail cyclization of ribosomally derived peptides promoted by aryl and alkyl amino-functionalized unnatural amino acids,”  Organic and Biomolecular Chemistry  2016145803-5812.
Tyagi V, Fasan* R  “Myoglobin-catalyzed olefination of aldehydes,”  Angewandte Chemie International Edition  201655(7)2512-2516.
Giovani S, Singh R, Fasan* R  “Efficient conversion of primary azides to aldehydes catalyzed by active site variants of myoglobin,”  Chemical Science  20167234-239.
Bionda N, Fasan* R  “Ribosomal synthesis of natural product-like bicyclic peptides in E. coli,”  ChemBioChem  201516(14)2011-2016.
Frost JR, Jacob NT, Papa LJ, Owens AE, Fasan* R  “Ribosomal Synthesis of Macrocyclic Peptides in Vitro and in Vivo Mediated by Genetically Encoded Amino-thiol Unnatural Amino Acids,”  ACS Chemical Biology  201510(8)1805-1816.
Tyagi V, Bonn RB, Fasan* R  “Intermolecular carbene S-H insertion catalysed by engineered myoglobin-based catalysts,”  Chemical Science  201562488-2494.
Smith JM, Fasan* R  “Synthesis of Macrocyclic Organo-Peptide Hybrids from Ribosomal Polypeptide Precursors via CuAAC-/hydrazide-mediated cyclization,”  Methods in Molecular Biology  2015128423-38.
Sreenilayam G, Fasan* R  “Myoglobin-catalyzed intermolecular carbene N-H insertion with arylamine substrates,”  Chemical Communications  2015511532-1534.
Bordeaux M, Tyagi V, Fasan* R  “Highly diastereo- and enantioselective olefin cyclopropanation via engineered myoglobin-based catalysts,”  Angewandte Chemie International Edition  201554(6)1744-1748.
Singh R, Kolev JN, Sutera PA, Fasan* R  “Enzymatic C(sp3)—H amination: P450-catalyzed conversion of carbonazidates into oxazolidinones>,”  ACS Catalysis  201551685-1691.
Bionda N, Cryan A, Fasan* R  “Bioinspired strategy for the ribosomal synthesis of thioether-bridged macrocyclic peptides in bacteria>,”  ACS Chemical Biology  20149(9)2008-2013.
Bordeaux M, Singh R, Fasan* R  “"Intramolecular C(sp3)—H amination of arylsulfonyl azides with engineered and artificial myoglobin-based catalysts>,”  Bioorganic and Medicinal Chemistry  201422(20)5697-5704.
Smith JM, Frost JR, Fasan R.  “Macrocyclic organo-peptide hybrids inhibit the interaction between p53 and HDM2/X by accommodating a functional α-helical motif,”  Chem. Comm.  201450(39)5027-5030.
Kolev JN, Zaengle JM, Ravikumar R, Fasan R.  “Enhancing the Efficiency and Regioselectivity of P450 Oxidation Catalysts via Unnatural Amino Acid Mutagenesis,”  Chembiochem.  201415(7)1001-1010.
Singh R, Bordeaux M, Fasan R.  “P450-catalyzed intramolecular sp3 C—H amination with arylsulfonyl azide substrates,”  ACS Catal.  20144(2)546-552.
Smith JM, Hill NC, Krasniak PJ, Fasan R.  “Synthesis of bicyclic organo-peptide hybrids via oxime/intein-mediated macrocyclization followed by disulfide bond formation,”  Org. Biomol. Chem.  201412(7)1135-1142.
Kolev JN, O'Dwyer KM, Jordan CT, Fasan R.  “Discovery of potent parthenolide-based antileukemic agents enabled by late-stage P450-mediated C―H functionalization,”  ACS Chem. Biol.  20149(1)164-173.
Smith JM, Frost JR, Fasan R.  “Emerging strategies to access peptide macrocycles from genetically encoded polypeptides,”  J. Org. Chem.  201378(8)3525-3531.
Frost JR, Vitali F, Jacob NT, Brown MD, Fasan R.  “Macrocyclization of organo-peptide hybrids via a dual bioorthogonal ligation; insights from structure-reactivity studies,”  Chembiochem  201314(1)146-160.
Zhang K, Shafer BM, Demars II MD, Stern HA, Fasan R.  “Controlled oxidation of remote sp3 C―H bonds in artemisinin via P450 catalysts with fine-tuned regio- and stereoselectivity,”  J. Am. Chem. Soc.  2012134(45)18695-18704.
Fasan, R.   “Tuning P450 Enzymes as Oxidation Catalysts,”  ACS Catal.  20122647-666.
Satyanarayana, M., Vitali, F., Frost, J. R., Fasan, R.   “Diverse organo-peptide macrocycles via a fast and catalyst-free oxime/intein-mediated dual ligation,”  Chem. Comm.  2012481461-1463.
Smith, J. M., Vitali, F., Archer, S. A., Fasan, R.  “Modular assembly of macrocyclic organo-peptide hybrids using synthetic and genetically encoded precursors,”  Angew. Chem. Int. Ed. Engl.  201150(22)5075-80.
Zhang, K., El Damaty, S., Fasan, R.  “P450 fingerprinting method for rapid discovery of terpene hydroxylating P450 catalysts with diversified regioselectivity,”  J. Am. Chem. Soc  2011133(10)3242-3245.
Rentmeister, A., Arnold, F. H., Fasan, R.  "Chemo-enzymatic fluorination of unactivated organic compounds,"  Nat. Chem. Biol.  2009526-28.
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Office: Hutchison 416
Phone: (585) 273-3504

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