Work

Protein Thioester Synthesis Enabled by Sortase

Year: 2012

Type: article

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 (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 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.

Source: Journal of the American Chemical Society

DOI: 10.1021/ja302354v

Authors: Jingjing J. Ling, Rocco L. Policarpo, Amy E. Rabideau, Xiaoli Liao, Bradley L. Pentelute

Institution: Massachusetts Institute of Technology

Language: English

Topic: Biochemical and Structural Characterization

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funder: National Institute of Allergy and Infectious Diseases

Awards: G421620279, New England Regional Center of Excellence in Biodefense and Emerging Infectious D