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Synthesis and screening of support-bound combinatorial cyclic peptide and free C-terminal peptide libraries

Joo, Sang Hoon
http://rave.ohiolink.edu/etdc/view?acc_num=osu1195561420

Abstract Details

2007, Doctor of Philosophy, Ohio State University, Chemistry.
One-bead one-compound (OBOC) peptide libraries have been useful tools in the biomedical sciences. However, OBOC peptide libraries usually display the N-termini of peptides on the surface as conventional solid phase peptide synthesis proceeds in the C to N direction. While large combinatorial libraries of cyclic peptides can be synthesized by the split-and-pool synthesis method, the sequence determination has been a challenge. Also, peptide libraries with free C-termini face the same problem as well as the difficulty of synthesis in the N to C direction. We report here the development of cyclic peptide libraries and C-terminal peptide libraries for high-throughput screening and sequencing. TentaGel microbeads (90 μm) were spatially segregated into outer and inner layers; cyclic peptides were displayed on the bead surface, whereas the inner core of each bead contained the corresponding linear encoding peptide. After screening of the cyclic peptide library, the identity of hit peptides was determined by sequencing the linear encoding peptides using a partial Edman degradation/mass spectrometry method. Using the same spatial segregation approach peptides were synthesized in the conventional C to N direction, with their C-termini attached to the support through an ester linkage on the bead surface but through an amide bond in the inner layer. The surface peptides were cyclized between N-terminal amine and a carboxyl group installed at a C-terminal linker sequence, while the internal peptides stayed in the linear form. Base hydrolysis of the ester linkage in the cyclic peptides exposed a free α-carboxyl group at the C-termini of the peptides attached to the resin via the N-termini. An inverted peptide library containing five random residues was synthesized and screened for binding to PDZ domains. The identity of the binding peptides was determined from the encoding peptides. Consensus recognition motifs were identified for the PDZ domains and representative peptides were individually synthesized and confirmed for binding to their cognate PDZ domains. These methods expanded the utility of OBOC peptide libraries by displaying peptides in different ways.
Dehua Pei (Advisor)
158 p.
Chemistry, Biochemistry
One-bead One-Compound (OBOC); partial Edman degradation; split-and-pool synthesis; cyclic peptides; Free C-terminal; peptide library; PDZ domain

Recommended Citations

Citations

  • Joo, S. H. (2007). Synthesis and screening of support-bound combinatorial cyclic peptide and free C-terminal peptide libraries [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1195561420

    APA Style (7th edition)

  • Joo, Sang Hoon. Synthesis and screening of support-bound combinatorial cyclic peptide and free C-terminal peptide libraries. 2007. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1195561420.

    MLA Style (8th edition)

  • Joo, Sang Hoon. "Synthesis and screening of support-bound combinatorial cyclic peptide and free C-terminal peptide libraries." Doctoral dissertation, Ohio State University, 2007. http://rave.ohiolink.edu/etdc/view?acc_num=osu1195561420

    Chicago Manual of Style (17th edition)

osu1195561420
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© 2007, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.