Fig. 1: Structural and functional characterization of human VLCAD by X-ray crystallography, HDX MS analysis, and liposomal translocation assay.
From: Structural basis for defective membrane targeting of mutant enzyme in human VLCAD deficiency
a Dimeric structure of human VLCAD ΔEx3 (PDB ID 7S7G) demonstrating its subcomponents and expanded MBR residues (purple) that extend outward from the protein core. See Supplementary Table 1 for data collection and results of refinement. b Crystal contacts between neighboring VLCAD dimers involve reciprocal interactions between the MBR of one dimer and the surface groove of another. c Relative deuterium uptake (%) of full-length VLCAD in solution after 10 s, 1 m, 10 m, and 1 h of deuteration. Residues are numbered using the mature (cleaved leader sequence) form of human VLCAD. HDX MS experiments were performed twice using independent preparations of VLCAD protein. See Supplementary Data 1 for the HDX MS data used to create this figure. d Translocation of VLCAD, but not its ΔMBR deletion mutant, to liposomes bearing the lipid composition of the inner mitochondrial membrane, as monitored by VLCAD western analysis of SEC fractions. The experiment was repeated twice with independent preparations of VLCAD protein with similar results. e Quantitation of VLCAD observed in liposomal translocation assay fractions (d) by densitometry using ImageJ software.
