Fig. 1: Nanobodies targeting protein palmitoylation.

A Schematic. Nanobody binding to its palmitoylated target protein delivers a thioesterase which removes the 16 carbon fatty acid palmitate (depicted in blue) from the target (Created in BioRender. Fulton, H. (2025) https://BioRender.com/n03e444). B Impact of anti-GFP nanobody LaG16 fused to the amino or carboxyl terminus of thioesterases (APT1, APT2) on YFP-Ca(v)1.2 I-II-linker palmitoylation. UF: unfractionated cell lysate; P: purified palmitoylated proteins; Flot2: flotillin-2. Data are means ± SEM from n = 8 independent experiments. C Impact of LaG16 fused to catalytically inactive (‘dead’) APTs on YFP-Ca(v)1.2 I-II-linker palmitoylation. Data are means ± SEM from n = 4 independent experiments. D Structure (PDB: 6LR7) of eGFP (green) in complex with LaG16 (magenta) highlighting the positions of F30, T55 and G103 (cyan) in CDRs 1, 2 and 3. E Wild-type and mutated (‘3W’) LaG16 binding to YFP-Ca(v)1.2 I-II-linker assessed by immunoprecipitating the FLAG tagged nanobody. This experiment was repeated independently three times with similar results. F Impact of LaG16 and mutated LaG16 fused to APTs on YFP-Ca(v)1.2 I-II-linker palmitoylation. Data are means ± SEM from n = 4 independent experiments. Statistical comparisons: one-way ANOVA followed by Tukey’s multiple comparisons test. Source data are provided as a Source Data file.