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. 2021 Oct 22;9:768854. doi: 10.3389/fchem.2021.768854

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Copyright © 2021 Chen, Zhang, Zhang, Wang, Liu and Tam.

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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Application of Vc1c in end-to-end cyclization of sfGFP. (A) Scheme of ligase-mediated sfGFP (generated from PDB code 2B3P) cyclization. sfGFP was co-expressed with an N-terminal Met-Ile and a C-terminal Asn-Ser-Leu-His6. The ligase can recognize and cleave the C-terminal motif Ser-Leu-His6 and perform end-to-end cyclization of sfGFP. (B) Upper: ESI-MS characterization of CM-sfGFP-NSL-His6 starting material or acyclic sfGFP (left) and cyclic sfGFP (right); Lower: Deconvolution of sfGFP cyclization (CM-sfGFP-NSL-His6: Cal. 28261, Obs. 28263; Cyclic sfGFP: Cal. 27220, Obs. 27220). The cyclization reaction was performed at pH 7.0, 37°C for 30 min using 25 µM CM-sfGFP-NSL-His6 and 500 nM Vc1c.