Fig. 5: The ZA domain binds weakly to the myrArf1:PH complex.
From: An active allosteric mechanism in ASAP1-mediated Arf1 GTP hydrolysis redefines PH domain function
A One-dimensional 1H NMR spectrum corresponding to the projection of an 1H-15N TROSY-HSQC along the F1 dimension obtained for ZA domain labeled PZA bound to ND alone (a) or bound to Arf (b), measured at the membrane surface of NDs containing PI(4,5)P2. The absence of visible changes indicates that the ZA domain reorients independently from the complex between Arf, PH and the ND, as expected from weak affinity between ZA and the ARF:PH complex. Motional reorientation is represented by a double-sided arrow. Isotopically labeled, NMR visible domain used in each experiment is colored in blue. B One-dimensional 1H NMR spectrum corresponding to the projection of an 1H-15N TROSY-HSQC along the F1 dimension obtained for myrArf1 bound to ND alone (a), in the presence of wt PH (b).The addition of a stoichiometric ratio of ASAP1-PH resulted in the loss of nearly all amide backbone resonances in the 1H-15N TROSY-HSQC spectrum, in stark contrast with the spectrum in the absence of the PH domain indicating that the G domain becomes locked, reorienting with the same correlation time as the nanodisc. Motional reorientation is represented by a double-sided arrow. Isotopically labeled, NMR visible domain used in each experiment is colored in blue. Source data are provided as a Source data file.
