Results: 4

1.
Figure 2

Figure 2. From: Role of Active Site Tyrosines in Dynamic Aspects of DNA Binding by AP endonuclease.

Single turnover binding of AP-site-containing DNA by AP endo mutants Y171F (▲) and Y171H (▼). For the experiment shown in the figure, substrate and enzyme were each present at 7 nM (Y171F) or 4 nM and 20 nM respectively (Y171H).

Luisa F. Melo, et al. DNA Repair (Amst). ;6(3):374-382.
2.
Figure 1

Figure 1. From: Role of Active Site Tyrosines in Dynamic Aspects of DNA Binding by AP endonuclease.

Single turnover binding of AP-site containing DNA by AP endonuclease mutants Y128A (●), Y269A (◆) and Y171A (□). For the experiment shown in the figure, substrate and enzyme were each present at 4 nM for Y128A, at 4 nM and 0.4 nM respectively for Y269A and at 7 nM for Y171A. [ES]eq for Y128A was 0.28 nM, when initial concentration of enzyme and substrate were each 4 nM. ([ES]eq for Y269A was 0.1 nM when initial enzyme and substrate were 0.4 nM and 4 nM respectively).

Luisa F. Melo, et al. DNA Repair (Amst). ;6(3):374-382.
3.

Figure 4. From: Role of Active Site Tyrosines in Dynamic Aspects of DNA Binding by AP endonuclease.

Comparison of binding AP-site containing DNA with (●, ◆) and without () an AP-site by EMSA analysis. WT and mutant proteins at the indicated concentrations were incubated with 0.25 nM substrate and resolved by non-denaturing gel electrophoresis as described in Methods. The distribution of bound and unbound substrate was quantitated by phosphorImager analysis.
A: Binding AP-site-containing DNA by Y128A(●) and Y269A (◆)
B: Binding by WT AP endo or Tyr171 mutants to oligonucleotide with (●) or without() an abasic site.

Luisa F. Melo, et al. DNA Repair (Amst). ;6(3):374-382.
4.
Figure 3

Figure 3. From: Role of Active Site Tyrosines in Dynamic Aspects of DNA Binding by AP endonuclease.

EMSA analysis of binding AP-site containing DNA by wild type and Y171A AP endonuclease, non-denaturing gel. WT or Y171A were incubated with 0.25 nM substrate for 30 min at 25 °C and resolved by non-denaturing gel electrophoresis as described in Methods. Lane 1, no added protein; lane 2, 0.5 nM WT; lane 3, 2.5 nM WT; lane 4, 5.0 nM WT; lane 5, 10 nM WT; lane 6, 25 nM WT; lane 7, 5 nM Y171A; lane 8, 25 nM Y171A; lane 9, 50 nM Y171A; lane 10, 100 nM Y171A; lane 11, 250 nM Y171A.

Luisa F. Melo, et al. DNA Repair (Amst). ;6(3):374-382.

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