Genetic PEGylation

Seiichi Tada,Takashi Andou,Yoshihiro Ito,Naoshi Dohmae,Takehiro Suzuki,Eiry Kobatake,Nediljko Budisa

doi:10.1371/journal.pone.0049235

Genetic PEGylation

Seiichi Tada, Takashi Andou + Show 5 more

Open Access

https://doi.org/10.1371/journal.pone.0049235

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Abstract

Polyethylene glycol (PEG) was genetically incorporated into a polypeptide. Stop-anticodon-containing tRNAs were acylated with PEG-containing amino acids and were then translated into polypeptides corresponding to DNA sequences containing the stop codons. The molecular weights of the PEG used were 170, 500, 700, 1000, and 2000 Da, and the translation was confirmed by mass spectrometry. The PEG incorporation ratio decreased as the molecular weight of PEG increased, and PEG with a molecular weight of 1000 Da was only slightly incorporated. Although improvement is required to increase the efficiency of the process, this study demonstrates the possibility of genetic PEGylation.

Highlights

Synthetic polymer–protein hybrids have been developed for use as therapeutic proteins or bioreactor enzymes [1,2,3,4,5,6,7,8,9,10]
The site-specific polymer attachment has been achieved with the complete synthetic construction of an erythropoietic protein [30,31], consisting of an a-amino acid polypeptide chain of 166 residues by using native chemical ligation
The method utilizes the UAG codon, which normally directs the termination of protein synthesis, to encode instead a nonnatural amino acid that is loaded onto the complementary tRNA

Summary

Introduction

Synthetic polymer–protein hybrids have been developed for use as therapeutic proteins or bioreactor enzymes [1,2,3,4,5,6,7,8,9,10]. The site-specific polymer attachment has been achieved with the complete synthetic construction of an erythropoietic protein [30,31], consisting of an a-amino acid polypeptide chain of 166 residues by using native chemical ligation. The method utilizes the UAG codon (the amber nonsense stop codon), which normally directs the termination of protein synthesis, to encode instead a nonnatural amino acid that is loaded onto the complementary tRNA.

Results

Conclusion