Abstract

This application requests support for the development of new methods for the incorporation of novel amino acids into recombinant proteins. Computational and experimental studies will be combined to design mutant forms of the aminoacyl-tRNA synthetases that are capable of efficient activation of amino acid analogs bearing reactive functional groups. Special attention will be given to amino acids that carry alkyne, azide and ketone functional groups, in order to provide powerful new chemistries for use in the engineering of natural and artificial proteins. Computational design will be evaluated in comparison with experimental approaches that involve multiple rounds of mutagenesis and selection, and computational predictions will be assessed by comparison with the results of in vitro kinetic studies of amino acid activation. Cell free transcription - translation systems will be used as appropriate to identify and address limitations due to poor amino acid transport. The new chemistries to be developed in this work will open new opportunities in the design of engineered biomaterials.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM062523-06
Application #
7283057
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Basavappa, Ravi
Project Start
2001-01-01
Project End
2009-08-31
Budget Start
2007-09-01
Budget End
2008-08-31
Support Year
6
Fiscal Year
2007
Total Cost
$323,949
Indirect Cost

  Institution

Name
California Institute of Technology
Department
Type
Schools of Engineering
DUNS #
009584210
City
Pasadena
State
CA
Country
United States
Zip Code
91125

  Publications

Stone, Shannon E; Glenn, Weslee S; Hamblin, Graham D et al. (2017) Cell-selective proteomics for biological discovery. Curr Opin Chem Biol 36:50-57
Mahdavi, Alborz; Hamblin, Graham D; Jindal, Granton A et al. (2016) Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis. J Am Chem Soc 138:4278-81
Bagert, John D; van Kessel, Julia C; Sweredoski, Michael J et al. (2016) Time-resolved proteomic analysis of quorum sensing in Vibrio harveyi. Chem Sci 7:1797-1806
Feng, Lihui; Rutherford, Steven T; Papenfort, Kai et al. (2015) A qrr noncoding RNA deploys four different regulatory mechanisms to optimize quorum-sensing dynamics. Cell 160:228-40
Yuet, Kai P; Doma, Meenakshi K; Ngo, John T et al. (2015) Cell-specific proteomic analysis in Caenorhabditis elegans. Proc Natl Acad Sci U S A 112:2705-10
Kwon, Inchan; Lim, Sung In (2015) Tailoring the substrate specificity of yeast phenylalanyl-tRNA synthetase toward a phenylalanine analog using multiple-site-specific incorporation. ACS Synth Biol 4:634-43
tom Dieck, Susanne; Kochen, Lisa; Hanus, Cyril et al. (2015) Direct visualization of newly synthesized target proteins in situ. Nat Methods 12:411-4
Van Deventer, James A; Yuet, Kai P; Yoo, Tae Hyeon et al. (2014) Cell surface display yields evolvable, clickable antibody fragments. Chembiochem 15:1777-81
Hatzenpichler, Roland; Scheller, Silvan; Tavormina, Patricia L et al. (2014) In situ visualization of newly synthesized proteins in environmental microbes using amino acid tagging and click chemistry. Environ Microbiol 16:2568-90
Yuet, Kai P; Tirrell, David A (2014) Chemical tools for temporally and spatially resolved mass spectrometry-based proteomics. Ann Biomed Eng 42:299-311

Showing the most recent 10 out of 61 publications