Work

Rapid Total Synthesis of DARPin pE59 and Barnase

Year: 2014

Type: article

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.

Source: ChemBioChem

DOI: 10.1002/cbic.201300797

Authors: Surin K. Mong, Alexander A. Vinogradov, Mark D. Simon, Bradley L. Pentelute

Institution: Massachusetts Institute of Technology

Language: English

Topic: Chemical Synthesis and Analysis

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funder: National Institute of General Medical Sciences

Award: Interdepartmental Biotechnology Training Program