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. Author manuscript; available in PMC: 2013 Mar 27.
Published in final edited form as: Biochemistry. 2012 Mar 16;51(12):2486–2495. doi: 10.1021/bi201704y

Figure 4.

Figure 4

Effect of TgPRF on formin-domain mediated TgACTI polymerization. (A) Comparison of polymerization kinetics of TgACTI in the presence or absence of formin domains with or without addition of equimolar TgPRF. Polymerization of TgACTI (5 μM) alone in F buffer alone (solid red), or TgACTI combined with 5 μM TgPRF (dashed red), 100 nM TgFRM1-FH1-FH2 (solid blue), 100 nM TgFRM1-FH1-FH2 and 5 μM TgPRF (dashed blue), 12 nM TgFRM2-FH1-FH2 (solid green), or 12 nM TgFRM2-FH1-FH2 and 5 μM TgPRF (dashed green) as monitored by light scattering. (B) Comparison of polymerization kinetics of TgACTI in the presence or absence of TgFRM1-FH1-FH2 and TgFRM2-FH1-FH2 in combination with and without addition of TgPRF. Polymerization of TgACTI (5 μM) in F buffer alone (solid red), or with 100 nM TgFRM1-FH1-FH2 (solid blue), 12 nM TgFRM2-FH1-FH2 (solid green), 100 nM TgFRM1-FH1-FH2 and 12 nM TgFRM2-FH1-FH2 (solid gray), or 100 nM TgFRM1-FH1-FH2, 12 nM TgFRM2-FH1-FH2 and 5 μM TgPRF (dashed gray) as monitored by light scattering. For comparison please note that the Y axes in A and B are different scales. (C) Comparison of polymerization kinetics of TgACTI in the presence or absence of formin with and without addition of 1:10 concentration of TgPRF. Polymerization of TgACTI (5 μM) in F buffer alone (solid red), or with 100 nM TgFRM1-FH1-FH2 (solid blue), 100 nM TgFRM1-FH1-FH2 and 0.5 μM TgPRF (dashed blue), 12 nM TgFRM2-FH1-FH2 (solid green), or with 12 nM TgFRM2-FH1-FH2 and 0.5 μM TgPRF (dashed green) as monitored by light scattering. Representative data from 2 or 3 experiments are shown. Purified recombinant TgPRF (after GST cleavage) was used for experiments shown.