Pairing this data with the known cancer-associated missense mutation frequency per residue (Fig. 1C), we selected 16 (17%) of the identified cancer-associated PBRM1-BD4 missense variants across 11 unique amino acid residues for recombinant expression and purification from Escherichia coli as isolated BD4 constructs, followed by in vitro biophysical characterization based on the criteria of observed in ≥3 patients, location at a conserved residue, or both (Table S1). The gene discussed is PBRM1; the disease is cancer.