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. 1998 Sep 29;95(20):11607–11612. doi: 10.1073/pnas.95.20.11607

Figure 1.

Figure 1

(A) Mutagenesis in the Walker A motifs of the p36, p37, and p40 subunits. The amino acid sequences of the Walker A motifs in the p36, p37, and p40 subunits are aligned. The conserved lysine residue (boxed) in each subunit was changed to alanine by site-directed mutagenesis. (B) Purification of wt and mutant p40·p37·p36 complexes. The purified wt and mutant p40·p37·p36 complexes (described in Materials and Methods) were analyzed by SDS/9% polyacrylamide gels. Lane 1, 3 μg of wt p40·p37·p36 complex; lanes 2–4, 3 μg of the mutant p40·p37·p36 complex with a mutation in p36 (p36K66A), p37 (p37K84A), and p40 (p40K82A), respectively. (C) ATPase activity of wt and mutant p40·p37·p36 complexes. ATPase measurements were carried out as described in Materials and Methods. Reaction mixtures contained wt and mutant p40·p37·p36 complexes in amounts as indicated, in the presence of 12.5 μM (as nucleotides) φX174 ssc DNA.