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.

Public Health Relevance

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.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM062159-14
Application #
9000702
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Gerratana, Barbara
Project Start
2001-02-01
Project End
2018-01-31
Budget Start
2016-02-01
Budget End
2017-01-31
Support Year
14
Fiscal Year
2016
Total Cost
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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