Incorporation of aliphatic proline residues into recombinantly produced insulin

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Date
2023
Authors
Breunig, Stephanie L.
Quijano, Janine C.
Donohue, Cecile
Henrickson, Amy
Demeler, Borries
Ku, Hsun Teresa
Tirrell, David A.
Journal Title
Journal ISSN
Volume Title
Publisher
American Chemical Society
Abstract
Analogs of proline can be used to expand the chemical space about the residue while maintaining its uniquely restricted conformational space. Here, we demonstrate the incorporation of 4R-methylproline, 4S-methylproline, and 4-methyleneproline into recombinant insulin expressed in Escherichia coli. These modified proline residues, introduced at position B28, change the biophysical properties of insulin: Incorporation of 4-methyleneproline at B28 accelerates fibril formation, while 4-methylation speeds dissociation from the pharmaceutically formulated hexamer. This work expands the scope of proline analogs amenable to incorporation into recombinant proteins and demonstrates how noncanonical amino acid mutagenesis can be used to engineer the therapeutically relevant properties of protein drugs.
Description
Open access article. Creative Commons Attribution 4.0 International license (CC BY 4.0) applies
Keywords
Dissociation , Genetics , Monomers , Nanofibers , Peptides and proteins , Prolines , Recombinant proteins , Insulin
Citation
Breunig, S. L., Quijano, J. C., Donohue, C., Henrickson, A., Demeler, B., Ku, H. T., & Tirrell, D. A. (2023). Incorporation of aliphatic proline residues into recombinantly produced insulin. ACS Chemical Biology, 18(12), 2574-2581. https://doi.org/10.1021/acschembio.3c00561
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