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Improved protein splicing using embedded split inteins.

ABSTRACT: Naturally split inteins mediate a traceless protein ligation process known as protein trans-splicing (PTS). Although frequently used in protein engineering applications, the efficiency of PTS can be reduced by the tendency of some split intein fusion constructs to aggregate; a consequence of the fragmented nature of the split intein itself or the polypeptide to which it is fused (the extein). Here, we report a strategy to help address this liability. This involves embedding the split intein within a protein sequence designed to stabilize either the intein fragment itself or the appended extein. We expect this approach to increase the scope of PTS-based protein engineering efforts.

SUBMITTER: Gramespacher JA

PROVIDER: S-EPMC5818749 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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Improved protein splicing using embedded split inteins.

Gramespacher Josef A JA Stevens Adam J AJ Thompson Robert E RE Muir Tom W TW

Protein science : a publication of the Protein Society 20180117 3

Naturally split inteins mediate a traceless protein ligation process known as protein trans-splicing (PTS). Although frequently used in protein engineering applications, the efficiency of PTS can be reduced by the tendency of some split intein fusion constructs to aggregate; a consequence of the fragmented nature of the split intein itself or the polypeptide to which it is fused (the extein). Here, we report a strategy to help address this liability. This involves embedding the split intein with ...[more]

PMID: 29226478

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