Fig. 1: Engineering of the duAb architecture.
From: Programmable protein stabilization with language model-derived peptide guides
A Building from prior work13, a YFP nanobody (YFP Nb) was linked to potent deubiquitinase catalytic domains using different linker candidates. Created in BioRender. Hong, L. (2025) https://BioRender.com/v65m595. B KCNQ1-YFP stabilization by YFP Nb-based stabilizers in HEK293T cells determined by flow cytometric analysis. Cells were co-transfected with a pcDNA3-Nedd4L vector in the presence or absence of 4 μM PR-619 DUB inhibitor as indicated. Data are the average of independent replicates (n = 3). L1 = GAPGSG, L2 = GSGSG. (C) KCNQ-YFP stabilization by YFP Nb-based stabilizers, specifically comparing the YFP Nb-L2-OTUB1 fusion with the OTUB1 C91S and OTUB1 D88A/C91S/H265A (ASA) mutants. Cells were co-transfected with a pcDNA3-Nedd4L vector. Data are the average of individual replicates (n = 3). For B, C, normalized cell fluorescence was calculated by dividing the percentage of YFP+ cells in a sample by that of (-) DUB with no DUB inhibitor for control cells. Statistical analysis was performed using the two-tailed Student’s t-test using GraphPad Prism 10 software, with calculated p values are represented as follows: *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****, p ≤ 0.0001. Samples with p value representations above their respective bars reflect comparisons between the control and that sample; all other p value notations compare those specific samples. Please refer to source data for numeric p values.
