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Incorporation of Aliphatic Proline Residues into Recombinantly Produced Insulin.

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.

SUBMITTER: Breunig SL

PROVIDER: S-EPMC10728891 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Incorporation of Aliphatic Proline Residues into Recombinantly Produced Insulin.

Breunig Stephanie L SL Quijano Janine C JC Donohue Cecile C Henrickson Amy A Demeler Borries B Ku Hsun Teresa HT Tirrell David A DA

ACS chemical biology 20231114 12

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, ...[more]

PMID: 37960878

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