Effects of Anti-R1 antibodies and R1 partial
peptides on SPB activation. (A) Anti-R1 antibodies inhibit SPB
activation. Affinity purified polyclonal antibodies against
R1—anti–RNR-N, anti–CH-MR, and anti-XRL522—and nonimmune rabbit IgG
were added separately to the SPB activator purified from
Xenopus egg mitotic extracts (Figure 1A, lanes f) to a
final concentration of 50–140 μg/ml, incubated on ice for 2 h,
and then subjected to the SPB activation assay. Horizontal axis, the
percentage of activated SPBs (N = 400). (B) Effects of R1 partial
peptide addition to the 200K fraction of Xenopus egg
mitotic extracts. The 200K fraction was mixed with each of the
peptides, RNR-N, PM1, PM1B, X384, M572, RNR-C, or M783, at 1.2 mM final
concentration andsubjected to the SPB activation
assay. Horizontal axis, the percentage of activated SPBs (N =
300). (C) The PM1 peptide suppresses SPB activation by the 200K
fraction in a concentration-dependent manner. The 200K fraction was
mixed with the PM1 peptide at various concentrations (0.17–1.3 mM) and
subjected to the SPB activation assay. Horizontal axis, the peptide
concentration (mM); vertical axis, the percentage of activated SPBs
(N = 100).