Journal: Nature Communications

Article Title: Structural basis for negative regulation of the Escherichia coli maltose system

doi: 10.1038/s41467-023-40447-y

Figure Lengend Snippet: a Affinity-purified MalT and MalY proteins were further analyzed by gel filtration using a Superdex 200 Increase 10/300 GL column in the presence of ADP (left). When MalT and MalY were incubated in a 1:1 molar ratio, the two proteins formed a stable complex with a molecular weight of about 300 kDa. The 9-mL peak observed when MalT is filtered alone may contain protein aggregates or MalT protein that partially oligomerized at a high protein concentration (>3 mg/ml). The presence of MalT and MalY in fractionated samples was confirmed by SDS-PAGE analyses (right). The experiments have been repeated for three times with similar results. b Cryo-EM density map with 2.94 Å resolution (top) and the refined structure model (middle) of the MalT-MalY complex (PDB: 8BOB) shown in different orientations. Each MalT or MalY protomer is labelled. Residue number and colors of each protein domain are indicated (bottom). The N-terminal segment of NBD is highlighted in yellow. The C-terminal domains of MalT are not well defined in the final structure. DBD DNA-binding domain.

Article Snippet: The experiments have been repeated for three times with similar results. b Cryo-EM density map with 2.94 Å resolution (top) and the refined structure model (middle) of the MalT-MalY complex (PDB: 8BOB) shown in different orientations.

Techniques: Affinity Purification, Filtration, Incubation, Molecular Weight, Protein Concentration, SDS Page, Cryo-EM Sample Prep, Binding Assay