This proposal is aimed at further developing and applying a general method that allows one to systematically add amino acids with novel physical, chemical and biological properties to the genetic codes of both prokaryotic and eukaryotic organisms. This methodology involves the generation of orthogonal tRNA/aminoacyl-tRNA synthetase (aaRS) pairs that uniquely recognize a noncoding codon and do not cross react with any of the endogenous tRNAs and aminoacyl-tRNA synthetases in the host organism. The amino acid specificity of the aminoacyl-tRNA synthetase is then modified such that it aminoacylates its cognate tRNA with only the desired unnatural amino acid and no endogenous amino acid. Using this method, a large number of structurally diverse unnatural amino acids have been incorporated efficiently and with high fidelity into proteins in response to nonsense and frameshift codons. We now propose to extend these studies in several directions. The first is focused on developing robust, user-friendly methods to genetically encode a large number of these unnatural amino acids in mammalian cells and Streptomyces. If successful this effort will significantly expand the scope and impact of this technology. The second effort focuses on the synthesis and in vitro and in vivo characterization of chemically defined antibody conjugates for the treatment of cardiovascular disease and acute myeloid leukemia. These latter efforts should not only pave the way for new or significantly improved therapeutics for major unmet medical needs, they should also provide a generalizable, straightforward approach to create similar classes of antibody conjugates for metabolic, cardiovascular and inflammatory diseases.
The ability to genetically encode unnatural amino acids beyond the common twenty will significantly enhance our ability to manipulate protein structure. The result will be new tools to study protein function in vitro and in vivo. It should also be possible to create homogenous 'chemically mutated' therapeutic proteins for cancer and cardiovascular disease with enhanced activity and improved pharmacology.
|Young, Douglas D; Schultz, Peter G (2018) Playing with the Molecules of Life. ACS Chem Biol 13:854-870|
|Xuan, Weimin; Collins, Daniel; Koh, Minseob et al. (2018) Site-Specific Incorporation of a Thioester Containing Amino Acid into Proteins. ACS Chem Biol 13:578-581|
|Xuan, Weimin; Shao, Sida; Schultz, Peter G (2017) Protein Crosslinking by Genetically Encoded Noncanonical Amino Acids with Reactive Aryl Carbamate Side Chains. Angew Chem Int Ed Engl 56:5096-5100|
|Xuan, Weimin; Schultz, Peter G (2017) A Strategy for Creating Organisms Dependent on Noncanonical Amino Acids. Angew Chem Int Ed Engl 56:9170-9173|
|Zambaldo, Claudio; Luo, Xiaozhou; Mehta, Angad P et al. (2017) Recombinant Macrocyclic Lanthipeptides Incorporating Non-Canonical Amino Acids. J Am Chem Soc 139:11646-11649|
|Xuan, Weimin; Yao, Anzhi; Schultz, Peter G (2017) Genetically Encoded Fluorescent Probe for Detecting Sirtuins in Living Cells. J Am Chem Soc 139:12350-12353|
|Luo, Xiaozhou; Fu, Guangsen; Wang, Rongsheng E et al. (2017) Genetically encoding phosphotyrosine and its nonhydrolyzable analog in bacteria. Nat Chem Biol 13:845-849|
|Mehta, Angad P; Li, Han; Reed, Sean A et al. (2016) Replacement of Thymidine by a Modified Base in the Escherichia coli Genome. J Am Chem Soc 138:7272-5|
|Luo, Xiaozhou; Zambaldo, Claudio; Liu, Tao et al. (2016) Recombinant thiopeptides containing noncanonical amino acids. Proc Natl Acad Sci U S A 113:3615-20|
|Liu, Yan; Wang, Ying; Zhang, Yong et al. (2016) Rational Design of Dual Agonist-Antibody Fusions as Long-acting Therapeutic Hormones. ACS Chem Biol 11:2991-2995|
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