Figure S3. Purification and cryo-EM... | Rockefeller University Press

Figure S3.

$Purification and cryo-EM analysis of the Vac17–Myo2 complex. (A) Example of preparative gel filtration showing co-expression of recombinant Vac17(H+MBD) and Myo2 tail. The black arrowhead indicates the complex formation for Vac17(H+MBD) and Myo2, which was sampled for data collection (top). The orange arrowhead marks free MBP, and the gray arrowhead indicates apo Myo2 tail. SDS-PAGE analysis of fractions from volumes 10–18 ml is shown. (B) Example micrograph with selected particles. Scale bar = 100 nm. (C) 2D class averages of particles processed using CryoSPARC. Scale bar = 130 Å. (D) Local resolution range of the reconstruction. (E) Model statistics (top) and Euler distribution of particles. (F) Comparison of the Myo2 tail (PDB accession no. 2F6H; yellow) with the AlphaFold model, trimmed to fit the cryo-EM reconstruction (gray). Black circles indicate regions resolved in the cryo-EM map. The red circle highlights a predicted region lacking empirical data. MBD, Myo2-binding domain. Source data are available for this figure: SourceData FS3.$

Purification and cryo-EM analysis of the Vac17–Myo2 complex. (A) Example of preparative gel filtration showing co-expression of recombinant Vac17(H+MBD) and Myo2 tail. The black arrowhead indicates the complex formation for Vac17(H+MBD) and Myo2, which was sampled for data collection (top). The orange arrowhead marks free MBP, and the gray arrowhead indicates apo Myo2 tail. SDS-PAGE analysis of fractions from volumes 10–18 ml is shown. (B) Example micrograph with selected particles. Scale bar = 100 nm. (C) 2D class averages of particles processed using CryoSPARC. Scale bar = 130 Å. (D) Local resolution range of the reconstruction. (E) Model statistics (top) and Euler distribution of particles. (F) Comparison of the Myo2 tail (PDB accession no. 2F6H; yellow) with the AlphaFold model, trimmed to fit the cryo-EM reconstruction (gray). Black circles indicate regions resolved in the cryo-EM map. The red circle highlights a predicted region lacking empirical data. MBD, Myo2-binding domain. Source data are available for this figure: SourceData FS3.

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