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Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing.

ABSTRACT: Manipulation of proteins by chemical modification is a powerful way to decipher their function. However, most ribosome-dependent and semi-synthetic methods have limitations in the number and type of modifications that can be introduced, especially in live cells. Here, we present an approach to incorporate single or multiple post-translational modifications or non-canonical amino acids into proteins expressed in eukaryotic cells. We insert synthetic peptides into GFP, NaV1.5 and P2X2 receptors via tandem protein trans-splicing using two orthogonal split intein pairs and validate our approach by investigating protein function. We anticipate the approach will overcome some drawbacks of existing protein enigineering methods.

SUBMITTER: Khoo KK

PROVIDER: S-EPMC7210297 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing.

Khoo K K KK Galleano I I Gasparri F F Wieneke R R Harms H H Poulsen M H MH Chua H C HC Wulf M M Tampé R R Pless S A SA

Nature communications 20200508 1

Manipulation of proteins by chemical modification is a powerful way to decipher their function. However, most ribosome-dependent and semi-synthetic methods have limitations in the number and type of modifications that can be introduced, especially in live cells. Here, we present an approach to incorporate single or multiple post-translational modifications or non-canonical amino acids into proteins expressed in eukaryotic cells. We insert synthetic peptides into GFP, Na<sub>V</sub>1.5 and P2X2 r ...[more]

PMID: 32385250

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