1H, 13C and 15N chemical shift assignments of the ZnR and GYF cytoplasmic domains of the GltJ protein from Myxococcus xanthus

Abstract

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A molecular switch controls assembly of bacterial focal adhesions

“…However, in the GltJ GYF domain (GYF GltJ ), as in nearly all other bacterial GYF domains, the G, Y, and F residues (after which the domain is named) are missing (fig. S2B) ( 24 ). We thus tested whether GYF GltJ interacts with the AglZ PRS sequence (residues 123 to 213; PRS AglZ ) using isothermal titration calorimetry (ITC) experiments.…”

“…NMR structure determination ZnR GltJ -and GYF GltJ -isolated domains have been produced as uniformly 13 C, 15 N-labeled proteins. Backbone and side-chain resonance assignments have been previously performed (24) and are available under Biological Magnetic Resonance Bank accession numbers 51104 and 51096 for ZnR GltJ and GYF GltJ , respectively. The 3D protein structures were calculated with the program CYANA using Nuclear Overhauser Effect (NOE) distance restraints obtained from Nuclear Overhauser Effect SpectroscopY (NOESY)-based NMR experiments and angular restraints (phi and psi) derived from chemical shifts of backbone atoms using the TALOS+ software program (37).…”

A novel molecular switch controls assembly of bacterial focal adhesions

“…Interestingly, GYF domains have been previously described as adaptor domains interacting with PRS through a conserved consensus signature 23 . However, in the GltJ GYF domain (GYF GltJ ), as in nearly all other bacterial GYF domains, the G,Y,F residues (after which the domain is named) are missing (Figure S3B) 24 . We thus tested whether GYF GltJ interacts with the AglZ PRS sequence (residues 123-213, PRS AglZ ) using Iso-Thermal Calorimetry (ITC) experiments.…”

“…ZnR GltJ and GYF GltJ isolated domains have been produced as uniformly 13 C, 15 Nlabeled proteins. Backbone and side-chains resonance assignments have been previously performed 24 and are available under BMRB accession number 51104 and 51096 for ZnR GltJ and GYF GltJ , respectively. The three-dimensional protein structures were calculated with the program CYANA using NOE distance restraints obtained from NOESY-based NMR experiments, and angular restraints (phi and psi) derived from chemical shifts of backbone atoms using the TALOS+ software program 39 .…”

“…ZnR GltJ (1-42) and GYF GltJ (138-222) isolated domains were expressed and purified as previously described (Attia et al, 2022). PRS AglZ (123-213), Nt1-222 (1-222) and linker-GYF GltJ were produced as N-terminal His6-tagged lipoyl fusion proteins.…”