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. Author manuscript; available in PMC: 2022 Dec 21.
Published in final edited form as: Bioconjug Chem. 2021 Oct 7;32(10):2233–2244. doi: 10.1021/acs.bioconjchem.1c00403

Figure 7:

Figure 7:

Non-templated native chemical ligation of E. coli 50S ribosomal subunit L32 (14a). (A) Sequence of L32 with L32-N (2-44, red) and L32-C (45-57, blue). The cleaved initiator Met was excluded. (B) Simplified schematic of the non-templated NCL between L32-N (12a) and L32-C (13a). Note that MTG was allowed to form the thioester for 10 min prior to TCEP addition, as shown in the more detailed scheme (Scheme S3). (C) HPLC chromatogram of the ligation endpoint. The ligation was considered complete after 48 h (after MTG addition), as nearly all of the reactive L32-N peptide (12a^) was depleted. However, a significant amount of L32-N peptide underwent hydrolysis (12a#) instead of ligation. 13a& refers to StBu-deprotected 13a. Analytical method B was used. HPLC and LC/MS chromatograms of more time points are shown in Fig. S49-50.