Computational design of a versatile, zero-radius proximity labeling enzyme

Supplementary Data 1. 2D Flow cytometry data for all LplA variants tested. Data are shown for (A) LplA orthologs, (B) ESM point mutants, (C) ProteinMPNN point mutants, (D) ESM combined mutants (E3-21), Directed evolution variant (D2), MPNN active-site redesigns (M1-24) and final combined engineering variants (F1-30). The black line in each plot shows the shape of the template (W37V LplA). Supplementary Data 2. Confocal microscopy of all ESM1B/1V predicted point mutations. mCherry-tagged LplA variants were imaged live in HEK 293T cells. All mutations were tested on the W37V-background as denoted by the superscript “V”. Supplementary Data 3. SEC-SAXS Elution profiles and Oligomer fits for LplA variants. Each document shows elution profile and associated SAXS scattering curves with Oligomer-predicted model fits. Data are plotted in both Log(I) vs q and Kratky plot format to better emphasize relevant differences, as well as the residual (data – model) for each fit. Supplementary Data 4. LplA structures used for SEC-SAXS Oligomer analysis. PDB files for all LplA structural models used for Oligomer analysis of the SAXS data. Supplementary Data 5. Code used to analyze SEC-SAXS data. Supplementary Data 6. Full-size/uncropped gels and blots. Uncropped scans of blots for main and extended data figures. (A) Uncropped scans of Figure 1F. (B) Uncropped scans of Figure 2A. (C) Uncropped scans of Figure 2C. (D) Uncropped scans of Figure 2D. (E) Uncropped scans of Figure 2E. (F) Uncropped scans of Figure 3E, and Extended Data Figure 7B, E. (G) Uncropped scans of Extended Data Figure 4A. (H) Uncropped scans of Extended Data Figure 4C. (I) Uncropped scans of Extended Data Figure 4E. (J) Uncropped scans of Extended Data Figure 5A-B. (K) Uncropped scans of Extended Data Figure 7G. (L) Uncropped scans of Extended Data Figure 7C-D. (M) Uncropped scans of Extended Data Figure8 B. (N) Uncropped scans of Extended Data Figure 8B. (O) Uncropped scans of Extended Data Figure 8C. (P) Uncropped scans of Extended Data Figure 8D (left). (Q) Uncropped scans of Extended Data Figure 8H. (R) Uncropped scans of Extended Data Figure 8D (right). S) Uncropped scans of Extended Data Figure 9B (left). (T) (Uncropped scans of Extended Data Figure 9B (right). (U) Uncropped scans of Extended Data Figure 10G.