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. 2022 Apr 8;9:857566. doi: 10.3389/fmolb.2022.857566

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Copyright © 2022 Prabhala, Gierach and Wood.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Ribbon diagram of the ΔI-CM mini-intein with an engineered N-terminal zinc binding motif and bound zinc ion interacting with a conserved His residue (ball-and-stick) close to the C-terminal cleaving junction. If properly designed, this motif will greatly increase the affinity for zinc by the intein, allowing a much higher sensitivity to zinc for suppressing cleaving during expression and purification of a fused target protein. The resulting intein is described in the associated patent (Ma et al., 2019; Patent No.: US 10323235 B2; prioritized in 2012, filed in 2017 and approved in 2019). The structure of PDB ID 2IN0 is here presented (Van Roey et al., 2007).