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. 2021 Dec 8;12:7137. doi: 10.1038/s41467-021-27184-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Overview of the structure of the complex where protein molecules are displayed in gray ribbon and MTX in ball and stick representations. The magenta ball represents a calcium ion bound to the nanoCLAMP. The loops of the nanoCLAMP randomized in the library are colored in green (V), magenta (W) and yellow (Z). The invariable loop 64–71 is colored in black. b Surface representation of the MTX:VHH complex with red colored structural elements in <3.2 Å proximity to the nanoCLAMP. In the figure, VHH is turned 90^o compared to (a). c superposition of the apo-VHH structure (PDB:3QXU) shown as gray ribbon with the structure of VHH:MTX complex (PDB:3QXV) displayed as blue ribbon and the structure of VHH:MTX from the ternary complex shown in red. Bound MTX molecules are displayed in the color of the respective complex. d Superposition of the apo form of the nanoCLAMP used a general library scaffold shown as gray ribbon with its structure from the nanoCLAMP from the ternary complex shown in blue with loops colored as in (a). e The omit map of MTX contoured at 3σ. f Graphic representation of the interface between the nanoCLAMP and VHH. The hydrogen bonds are shown as blue dashed lines and their lengths are displayed.