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Enzymatic Construction of Protein Polymer/Polyprotein Using OaAEP1 and TEV Protease.

ABSTRACT: The development of chemical and biological coupling technologies in recent years has made possible of protein polymers engineering. We have developed an enzymatic method for building polyproteins using a protein ligase OaAEP1 (asparagine endopeptidase 1) and protease TEV (tobacco etching virus). Using a mobile TEV protease site compatible with the OaAEP1 ligation, we achieved a stepwise polymerization of the protein on the surface. The produced polyprotein can be verified by protein unfolding scenario using atomic force microscopy-based single-molecule force spectroscopy (AFM-SMFS). Thus, this study provides an alternative method for polyprotein engineering and immobilization.

SUBMITTER: Deng Y

PROVIDER: S-EPMC7842765 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Enzymatic Construction of Protein Polymer/Polyprotein Using OaAEP1 and TEV Protease.

Deng Yibing Y Shi Shengchao S Zheng Bin B Wu Tao T Zheng Peng P

Bio-protocol 20200420 8

The development of chemical and biological coupling technologies in recent years has made possible of protein polymers engineering. We have developed an enzymatic method for building polyproteins using a protein ligase OaAEP1 (asparagine endopeptidase 1) and protease TEV (tobacco etching virus). Using a mobile TEV protease site compatible with the OaAEP1 ligation, we achieved a stepwise polymerization of the protein on the surface. The produced polyprotein can be verified by protein unfolding sc ...[more]

PMID: 33659562

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