Studies of the interaction between RFC and PCNA. (A)
Surface plasmon resonance analyses. Indicated concentrations of RFC
were injected into a sensor chip prefixed with about 1,000 resonance
units of PCNA. All of the procedures were performed with 150 mM NaCl
Running Buffer, with RFC as the analyte without nucleotides (1) or with
1 mM ATP (2), ATPγS (3), or ADP (4). KD
values with SD obtained from three injections were (1 ± 0.4)
× 10−9 M (2 ± 0.5) × 10−10 M
(9 ± 2) × 10−10 M and (1 ± 0.8) ×
10−8 M, respectively. (B) Gel mobility
profiles of RFC and RFC-PCNA complexes without ATP (−, lanes 2 and 7),
with ATP (lanes 3 and 8), with ADP (lanes 4 and 9), or with ATPγS
(lanes 5 and 10) at 0.15 M (lanes 1–5) or 0.5 M (lanes 6–10) NaCl.
Arrows indicate migration positions of silver-stained RFC and RFC-PCNA
complex. In lanes 1 and 6, only RFC was applied. PCNA runs at the dye
front and does not appear in this picture under these electrophoresis
conditions.