Specific Disulfide Bond Formation Directed by Peptide Nucleic Acids

Lin Cai,Wen Hou

doi:10.21577/0103-5053.20190263

Abstract

In this study, we successfully demonstrated a strategy for specific disulfide bond formation by hybridization of two complementary peptide nucleic acid strands. Peptides were linked to the pair of complementary peptide nucleic acids by native chemical ligation, respectively, and base pairing drove the two peptides into close proximity for interchain disulfide bond formation. Since peptide nucleic acids directed system is easier to manipulate than deoxyribonucleic acid system, this system will be more useful and have a broader application scope in the construction of complex biomolecular structures.

Highlights

Peptide nucleic acid (PNA) is an oligonucleotide mimic in which theribose-phosphate backbone has been replaced by an N-(2-aminoethyl)glycine unit and the nucleobases (adenyl (A), cytosyl (C), guanidyl (G), and thymidyl (T)) are attached to the backbone through methylenecarbonyl linkages (Figure 1).[1]
We report a system for peptide heterodimerization through PNA-templated disulfide bond formation (Figure 2)
PNA was successfully synthesized by Fmoc solid phase peptide synthesis (SPPS) protocol with minor modifications

Summary

Introduction

Peptide nucleic acid (PNA) is an oligonucleotide mimic in which the (deoxy)ribose-phosphate backbone has been replaced by an N-(2-aminoethyl)glycine unit and the nucleobases (adenyl (A), cytosyl (C), guanidyl (G), and thymidyl (T)) are attached to the backbone through methylenecarbonyl linkages (Figure 1).[1]. The PNA residues are linked by amide bond like peptide and PNA has the hybridization properties of nucleic acids originally detailed by Watson and Crick in 1953.2 Since the backbone of PNA does not contain charged phosphate groups, the complementary PNA/PNA strands or PNA/DNA or PNA/RNA (ribonucleic acid) strands are more stable and have higher melting temperature (Tm) than the Figure 1. Our strategies are utilizing the hybridization properties of two complementary strands of PNA to bring together two peptides or proteins attached to each strand to approach some biological or chemical activities, such as directed disulfide bonds formation. This idea was originally inspired by the DNA-directed assembly of enzyme complexes, which was reviewed below.

Methods

Results and Discussion

Conclusions