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. Author manuscript; available in PMC: 2016 Dec 24.
Published in final edited form as: Methods Enzymol. 2016 Jun 24;580:223–250. doi: 10.1016/bs.mie.2016.05.009

Figure 1.

Figure 1

Scheme showing the iterative engineering of a soluble, non-self-associating protein building block by MDPSA and MeTIR. By MDPSA, chelating motifs (e.g., bis-His clamps) can be incorporated into the protein surface, enabling the formation of metal-directed oligomers. By MeTIR, the nascent interfaces in these oligomers can be reinforced with the installation of complementary PPIs to generate self-assembling complexes that can form in the absence of templating metals. The installation of disulfide crosslinks increases scaffold robustness and the fidelity of self-assembly. These robust scaffolds can accommodate reactive metal sites to generate de novo designed metalloenzymes.