Protein Thioester Synthesis Enabled by Sortase

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 recomb...

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Bibliographic Details
Published in:Journal of the American Chemical Society 2012-07, Vol.134 (26), p.10749-10752
Main Authors: Ling, Jingjing J., Policarpo, Rocco L., Rabideau, Amy E., Liao, Xiaoli, Pentelute, Bradley L.
Format: Article
Language:English
Subjects:
Aminoacyltransferases - metabolism
Antigens, Bacterial - chemistry
Bacterial Proteins - metabolism
Bacterial Toxins - chemistry
Cysteine Endopeptidases - metabolism
Esters - chemistry
Inteins
Peptides - chemical synthesis
Protein Transport
Proteins - chemical synthesis
Recombinant Proteins - chemistry
Sulfhydryl Compounds - chemistry
Sulfur Compounds
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Summary: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 αthioester. We used our method to prepare two different anthrax toxin cargo proteins: one containing an αthioester 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.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja302354v