Figure 7.
Computational modeling predicts a basic amphipathic α-helix motif in the C-terminus anchors Rad to the plasma membrane. (A) AlphaFold prediction of M. musculus Rad full-length (Uniprot accession no. O88667) was input to the PPM 3.0 web server tool for computational modeling of Rad with a mammalian membrane. The inset image focuses on the predicted embedded residues that are amino acids in the predicted helix 8 (H8) and highlights the alanine which was replaced with a stop codon in the Flag-RadΔCT mouse model. AlphaFold was then used to predict a structure for RadΔCT. Modelling RadΔCT (1–276) with PPM 3.0 predicts low probability of membrane interaction. (B) Abbreviated protein schematic of full length Rad and RadΔCT. The GTPase core is shown with its putative interaction with the L-type calcium channel CaVβ subunit. The amino acids of the predicted amphipathic α-helix basic residues are highlighted in green. Helices 7 (black) and 8 (green) correspond to the colored H7 and H8 in the Rad proteins in A. (C) Tabular results of PPM 3.0 predictions. Full-length Rad ΔGtransfer was increased 160% relative to RadΔCT. The RadΔCT ΔGtransfer of −2.8 kcal/mol is relatively low, suggesting a low probability for hydrophobic surfaces involved in protein–membrane interactions. The predicted residues (152 and 154) for RadΔCT reside in the GTPase core that is known to interact with CaVβ.

Computational modeling predicts a basic amphipathic α-helix motif in the C-terminus anchors Rad to the plasma membrane. (A) AlphaFold prediction of M. musculus Rad full-length (Uniprot accession no. O88667) was input to the PPM 3.0 web server tool for computational modeling of Rad with a mammalian membrane. The inset image focuses on the predicted embedded residues that are amino acids in the predicted helix 8 (H8) and highlights the alanine which was replaced with a stop codon in the Flag-RadΔCT mouse model. AlphaFold was then used to predict a structure for RadΔCT. Modelling RadΔCT (1–276) with PPM 3.0 predicts low probability of membrane interaction. (B) Abbreviated protein schematic of full length Rad and RadΔCT. The GTPase core is shown with its putative interaction with the L-type calcium channel CaVβ subunit. The amino acids of the predicted amphipathic α-helix basic residues are highlighted in green. Helices 7 (black) and 8 (green) correspond to the colored H7 and H8 in the Rad proteins in A. (C) Tabular results of PPM 3.0 predictions. Full-length Rad ΔGtransfer was increased 160% relative to RadΔCT. The RadΔCT ΔGtransfer of −2.8 kcal/mol is relatively low, suggesting a low probability for hydrophobic surfaces involved in protein–membrane interactions. The predicted residues (152 and 154) for RadΔCT reside in the GTPase core that is known to interact with CaVβ.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal