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Protein thioester synthesis enabled by sortase.

ABSTRACT: Proteins containing a C-terminal thioester are important intermediates in semisynthesis. Currently there is one main method for the synthesis of protein thioesters that relies upon the use of engineered inteins. Here we report a simple strategy, utilizing sortase A, for routine preparation of recombinant proteins containing a C-terminal (?)thioester. We used our method to prepare two different anthrax toxin cargo proteins: one containing an (?)thioester and another containing a D-polypeptide segment situated between two protein domains. We show that both variants can translocate through protective antigen pore. This new method to synthesize a protein thioester allows for interfacing of sortase-mediated ligation and native chemical ligation.

SUBMITTER: Ling JJ

PROVIDER: S-EPMC3465687 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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Protein thioester synthesis enabled by sortase.

Ling Jingjing J JJ Policarpo Rocco L RL Rabideau Amy E AE Liao Xiaoli X Pentelute Bradley L BL

Journal of the American Chemical Society 20120619 26

Proteins containing a C-terminal thioester are important intermediates in semisynthesis. Currently there is one main method for the synthesis of protein thioesters that relies upon the use of engineered inteins. Here we report a simple strategy, utilizing sortase A, for routine preparation of recombinant proteins containing a C-terminal (α)thioester. We used our method to prepare two different anthrax toxin cargo proteins: one containing an (α)thioester and another containing a D-polypeptide seg ...[more]

PMID: 22686546

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