Journal Article

Characterization and optimization of two-chain folding pathways of insulin via native chain assembly

Hojo, Hironobu, Noguchi, Masato, Katayama, Hidekazu, Okumura, Masaki, Inaba, Kenji, Arai, Kenta, Takei, Toshiki, Shinozaki, Reina, Ando, Setsuko, Iwaoka, Michio, Fujisawa, Shouta, Moroder, Luis

2018

Overview

Until recently the total synthesis of insulin, with its characteristic heterodimeric structure crosslinked by two interchain and one intrachain disulfide (SS) bridge, remained largely an unsolved challenge. By optimizing the synthesis and directed disulfide crosslinking of the two chains, and by applying biomimetic monocomponent proinsulin approaches, efficient insulin syntheses have been realized. Here we report the optimization and characterisation of an alternative strategy, oxidative native chain assembly. In this method unprotected A- and B-chains assemble oxidatively under thermodynamic control to afford bovine pancreatic insulin in 39% yield. Folding is found to proceed predominantly via structured 1SS* and 2SS* intermediates with a common interchain Cys A20 ‒Cys B19 disulfide. These results suggest that native chain assembly, long considered inefficient, may represent a reasonable strategy to access insulin variants. This is supported by the synthesis of human insulin and human type-II relaxin in yields of up to 49 and 47%, respectively, although the application to human insulin Val A16 variant is unsuccessful. The synthesis and folding pathways of insulin and related proteins are of wide interest. Here the authors characterise the major two-chain oxidative folding pathways of bovine pancreatic insulin, and develop synthetic conditions applicable to related foldable insulin variants

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/45/776/1178 / 631/92/611 / 639/638/92/470 / 82/16 / Assembly / Biomimetics / Chains / Chemistry / Chemistry and Materials Science / Chemistry/Food Science / Crosslinking / Folding / Insulin / Optimization / Synthesis