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. 2017 Mar 10;8:14671. doi: 10.1038/ncomms14671

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Copyright © 2017, The Author(s)

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Proteins secreted into the culture medium after site-directed mutagenesis of the residues indicated were detected by western blotting using the N-terminal His-tag present on either or both the α1(I) and α2(I) chains, as indicated by H (for example, A1H means His-tagged α1 chain). The α2(I) chain is indicated by the green/blue colour. Gels were run in non-reducing conditions, where trimers migrate at 85 kDa and monomers at 30 kDa. (a) Controls and mutants using the His-tagged α1(I) chain. (b) Controls and mutants using the non-tagged α1(I) chain and the His-tagged α2(I) chain. (c) Schematic view down the axis of a C-propeptide heterotrimer showing interacting residues at the homo- and hetero-chain interfaces. Same colour coding throughout. Data shown are representative of triplicate (a) or duplicate (b) biological replicates.