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Rapid total synthesis of DARPin pE59 and barnase.

ABSTRACT: We report the convergent total synthesis of two proteins: DARPin pE59 and Bacillus amyloliquefaciens RNase (Barnase). Leveraging our recently developed fast-flow peptide-synthesis platform, we rapidly explored numerous conditions for the assembly of long polypeptides, and were able to mitigate common side reactions, including deletion and aspartimide products. We report general strategies for improving the synthetic quality of difficult peptide sequences with our system. High-quality protein fragments produced under optimal synthetic conditions were subjected to convergent native chemical ligation, which afforded native full-length proteins after a final desulfurization step. Both DARPin and Barnase were folded and found to be as active as their recombinant analogues.

SUBMITTER: Mong SK

PROVIDER: S-EPMC4045705 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature

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Rapid total synthesis of DARPin pE59 and barnase.

Mong Surin K SK Vinogradov Alexander A AA Simon Mark D MD Pentelute Bradley L BL

Chembiochem : a European journal of chemical biology 20140311 5

We report the convergent total synthesis of two proteins: DARPin pE59 and Bacillus amyloliquefaciens RNase (Barnase). Leveraging our recently developed fast-flow peptide-synthesis platform, we rapidly explored numerous conditions for the assembly of long polypeptides, and were able to mitigate common side reactions, including deletion and aspartimide products. We report general strategies for improving the synthetic quality of difficult peptide sequences with our system. High-quality protein fra ...[more]

PMID: 24616257

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