Selected regions of 13C-1H HMQC spectra of (A) wt and (B) R95G [2H,12C, proR ILVM 13CH3]-labeled ND1Lp97-ADP, 800 MHz, 50°C with and without 1.5 fold excess p47 UBX domain. Assignment of methyl groups is provided for residues involved in UBX binding in orange, as reported in a crystallographic study (Dreveny et al., 2004). Assignments in black indicate residues that respond to the NTD up/down equilibrium shift, which is not affected by binding of the p47 UBX domain. Orange arrows connect peaks that change in position upon binding the UBX domain. (C) Binding of full-length p47 trimer to wt and R95G ND1Lp97-ADP (1.2 fold excess). Residues with CSPs in response to changes to the NTD up/down equilibrium have been selected; note the difference in positions between peaks in red (corresponding to wt ND1Lp97-ADP) and peaks in blue (R95G ND1Lp97-ADP) the reflects differences in pU between wt and R95G p97. Binding of p47 does not alter the up/down equilibrium, as many of the residues that are reporters of the equilibrium (via changes in peak positions upon mutation) do not have CSPs upon p47 adaptor binding (compare red and grey for wt; blue and black for R95G and methyl probes from V99, V201, L229, M427, M442). Alternatively, in the case where binding does cause CSPs (L140, I119), these are not in a direction that indicates changes to the up/down equilibrium.