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. 2016 Nov 18;7:13376. doi: 10.1038/ncomms13376

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Copyright © 2016, The Author(s)

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(a) Amino acid differences among functional alleles of IGHV1-69. (b) The VH of clone D2-06-N2 was germline-reverted to all alleles with amino acid differences, and tested for binding to IsdB by ELISA. All alleles with a G50R substitution lost binding. ELISA data is an average of three independent experiments. Error bars are defined as s.d. (c.) Three individual variants (G50R, F54L and T56I) of D2-06-N2 (IGHV1-69*01 germline-reverted) were generated and their binding to IsdB was tested. Only variant G50R showed significant loss of binding. ELISA data is an average of three independent experiments. Error bars are defined as s.d. (d) Analysis of the structure illustrates how a change from G to R (most frequent rotamer) at position 50 is expected to cause a steric clash in the binding to NEAT2.