Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Nov 20;14:7551. doi: 10.1038/s41467-023-43148-8

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 3 — a Liposome adhesion assay using extruded liposomes coated with His-tagged zGFRα1 constructs as indicated. Liposome clustering was monitored by taking time-course measurements of absorbance at 650 nm (OD650). Arrow shows timepoint at which soluble zGDNF^mat was added. Error bars represent standard deviation from three technical repeats. b 2D tomographic slices from reconstructed tomograms of zGDNF^mat-zGFRα1^D2-D3+-mediated liposome aggregates. Images show close-up views of bridging protein density between two liposome membranes (indicated with white arrow heads). Scale bar: 20 nm. c Subtomogram map of the zGDNF^mat-zGFRα1^D2-D3+ adhesion complex bridging adhered liposomes with the decameric complex crystal structure fitted into the map using ChimeraX⁷⁷ ‘fit-in-map’ tool. Orthongonal views are shown. d Structural overlap between the high affinity GFRα1–RET interface and the GFRα1 pentameric interface. Top right, view of zGDNF-zGFRα1-zRET^ECM complex (PDB: 7AML) projecting down the two-fold molecular dyad. One zGFRα1 protomer is represented as a surface rendering coloured according to domain, zGFRα1-D1 light green, zGFRα1-D2 green and zGFRα1-D3 dark green. Top left, close up view of zRET^ECM and zGFRα1 interaction at a calcium-junction-D3 domain interface. Selected interacting residues shown as sticks and calcium atoms shown as blue spheres. Bottom right, view of the pentameric interface from the decameric zGDNF^mat-zGFRα1^D2-D3+ structure highlighting a single zGFRα1 protomer by surface rendering. Bottom left, close-up view of the pentameric interface highlighting the same interacting residues as shown for RET interaction. e Schematic representation of the LiCAM capacity of GFRα1 under defined conditions. (i) zGFRα1^D2-D3+ acts as a strong adhesion molecule upon zGDNF^mat addition through the ability to form homophilic interactions in cis. (ii) R170E mutation in zGFRα1^D2-D3+ targets the ability of zGFRα1 to form interactions in trans and thereby abolishes the adhesive capacity of zGFRα1. (iii) Mutations targeting the cis pentamer interface, K254E L290E, lead to reduced adhesion function by zGFRα1 in the presence of zGDNF^mat. (iv) In the presence of zRET^ECM, zGFRα1^D2-D3+ preferentially forms a ternary complex in cis upon zGDNF^mat addition.