Table 2.
Structure-based mutagenesis of HopA1
a
Complex formation was assayed as described in Materials and Methods on the basis of recombinant bacterial coexpression and nickel affinity pull-down of untagged full-length GST-tagged HopA1 with dodecahistidine-tagged ShcA chaperone. This purification scheme was used to isolate milligram quantities of the complex used in crystallization studies. +, robust stable complex formation in affinity pull-down and size exclusion chromatography analyses; +/−, a significantly destabilized complex in all assays; −, a stable complex could not be isolated.
b
Residue in the β-motif.