Sequence-Defined Scaffolding of Peptides on Nucleic Acid Polymers.
We have developed a method for the T4 DNA ligase-catalyzed DNA-templated polymerization of 5'-phosphorylated pentanucleotides containing peptide fragments. The polymerization proceeds sequence-specifically to generate DNA-scaffolded peptides in excellent yields. The method has been shown to tolerate peptides ranging from two to eight amino acids in length with a wide variety of functionality. We validated the capabilities of this system in a mock selection for the enrichment of a His-tagged DNA-scaffolded peptide phenotype from a library, which exhibited a 190-fold enrichment after one round of selection. This strategy demonstrates a promising new approach to allowing the generation and in vitro selection of high-affinity reagents based upon single-stranded DNA scaffolding of peptide fragments.
Department of Chemistry, University of Georgia , 140 Cedar Street, Athens, Georgia 30602-2556, United States.
Chun
C
Guo
Department of Chemistry, University of Georgia , 140 Cedar Street, Athens, Georgia 30602-2556, United States.
Christopher P
CP
Watkins
Department of Chemistry, University of Georgia , 140 Cedar Street, Athens, Georgia 30602-2556, United States.
Ryan
R
Hili
D055162:Biocatalysis; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism*; D015394:Molecular Structure; D009696:Nucleic Acids / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D058105:Polymerization; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*
D009696:Nucleic Acids; D010455:Peptides; D011108:Polymers; D004247:DNA; D011088:DNA Ligases