Figure 7. The CH of ATL2-1 directly... | Rockefeller University Press

The CH of ATL2-1 directly interacts with the GTPase domain of ATL2-1. (A) His6-tagged ATL2 (1–480) was mixed with the ATL2-1 peptide, the ATL2-1 (KKE) peptide, or the ATL2-2 peptide at 4°C for 1 h. The protein–peptide mixtures were then further incubated with Ni-NTA agarose (Qiagen) at 4°C for 30 min. Proteins and peptides bound to Ni-NTA agarose were analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining. (B) His6-tagged ATL2 (1–480), his6-tagged ATL2 (1–373), or his6-tagged ATL2 (373–480) was mixed with the ATL2-1 peptide at 4°C for 1 h. The protein–peptide mixtures were then further incubated with Ni-NTA agarose (Qiagen) at 4°C for 30 min. Proteins and peptides bound to Ni-NTA agarose were analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining. (C) AlphaFold structure prediction of ATL2-1 rendered in cartoon form in PyMOL. The per-residue confidence score (pLDDT) and predicted aligned error (PAE) of this predicted structure are shown in Fig. S4. Source data are available for this figure: SourceData F7.

Figure 7.

The CH of ATL2-1 directly interacts with the GTPase domain of ATL2-1. (A) His₆-tagged ATL2 (1–480) was mixed with the ATL2-1 peptide, the ATL2-1 (KKE) peptide, or the ATL2-2 peptide at 4°C for 1 h. The protein–peptide mixtures were then further incubated with Ni-NTA agarose (Qiagen) at 4°C for 30 min. Proteins and peptides bound to Ni-NTA agarose were analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining. (B) His₆-tagged ATL2 (1–480), his₆-tagged ATL2 (1–373), or his₆-tagged ATL2 (373–480) was mixed with the ATL2-1 peptide at 4°C for 1 h. The protein–peptide mixtures were then further incubated with Ni-NTA agarose (Qiagen) at 4°C for 30 min. Proteins and peptides bound to Ni-NTA agarose were analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining. (C) AlphaFold structure prediction of ATL2-1 rendered in cartoon form in PyMOL. The per-residue confidence score (pLDDT) and predicted aligned error (PAE) of this predicted structure are shown in Fig. S4. Source data are available for this figure: SourceData F7.

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