CaltechTHESIS
  A Caltech Library Service

N-terminal modification and codon reassignment with non-canonical amino acids in proteins

Citation

Connor, Rebecca E. (2008) N-terminal modification and codon reassignment with non-canonical amino acids in proteins. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechETD:etd-03052008-065324

Abstract

Proteins are ubiquitous macromolecules that effect and control all the processes of life from reproduction to respiration to physical motion. These diverse molecules also provide physical structure and defensive mechanisms. The twenty canonical amino acids can be found in virtually every protein; however, in some organisms, the set of endogenous amino acids also contains residues outside the “canon,” such as pyrrolysine, selenocysteine, and formylmethionine. Although a range of chemistries is available through natural side-chain diversity, some functionalities such as halogens, ketones, azides, alkenes, and alkynes are not found in nature. The introduction of a broader range of chemical functionality into proteins and protein-based materials through the use of non-canonical amino acids represents a challenging goal for protein engineering. The persistence of all the amino acids throughout protein sequences also presents a challenge for biochemical modification at a particular location. The insertion of a non-natural amino acid at a single location on a protein can allow specific modification without further affecting the natural protein sequence. The focus of this thesis is on a new method for the post-translational site-specific introduction of non-canonical amino acids to the N-terminus of proteins in vitro and progress made towards developing a complementary in vivo method using the E. coli L,F-transferase. Addition of non-proteinogenic functionality to the coat proteins of M13 bacteriophage using non-canonical amino acids is also explored.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:bioconjugates; non-canonical amino acids; protein engineering; protein modification; transferases
Degree Grantor:California Institute of Technology
Division:Chemistry and Chemical Engineering
Major Option:Chemistry
Thesis Availability:Mixed availability, specified at file level
Research Advisor(s):
  • Tirrell, David A.
Thesis Committee:
  • Barton, Jacqueline K. (chair)
  • Varshavsky, Alexander J.
  • Tirrell, David A.
  • Shan, Shu-ou
Defense Date:19 February 2008
Record Number:CaltechETD:etd-03052008-065324
Persistent URL:http://resolver.caltech.edu/CaltechETD:etd-03052008-065324
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:878
Collection:CaltechTHESIS
Deposited By: Imported from ETD-db
Deposited On:06 Mar 2008
Last Modified:26 Dec 2012 02:33

Thesis Files

[img] PDF (Appendix1.pdf) - Final Version
Restricted to Caltech community only
See Usage Policy.

1561Kb
[img]
Preview
PDF (ConnorChapter1.pdf) - Final Version
See Usage Policy.

594Kb
[img] PDF (ConnorChapter2.pdf) - Final Version
Restricted to Caltech community only
See Usage Policy.

1026Kb
[img]
Preview
PDF (ConnorChapter3.pdf) - Final Version
See Usage Policy.

1173Kb
[img]
Preview
PDF (ConnorTOC.pdf) - Final Version
See Usage Policy.

150Kb

Repository Staff Only: item control page