Table 1.
TR-F and cosedimentation binding parameters of cMyBP-C C0–C2 binding to actin and actin–Tm
TR-FCosedimentation
FilamentPKAKd (μM)Bmax (maximal Δτ)Kd (μM)Bmax (fraction bound; C0–C2/actin)
Actin − 2.5 ± 0.4 5.1 ± 0.3 4.3 ± 0.3 1.56 ± 0.04 
Actin 19 ± 5a 9 ± 1a 6.5 ± 0.3a 1.48 ± 0.03 
Actin–Tm − 1.4 ± 0.5b 4.6 ± 0.4 3.3 ± 0.4 1.61 ± 0.06 
Actin–Tm 3.6 ± 0.7a,c 4.5 ± 0.3c 6.6 ± 0.4a 1.46 ± 0.04 
TR-FCosedimentation
FilamentPKAKd (μM)Bmax (maximal Δτ)Kd (μM)Bmax (fraction bound; C0–C2/actin)
Actin − 2.5 ± 0.4 5.1 ± 0.3 4.3 ± 0.3 1.56 ± 0.04 
Actin 19 ± 5a 9 ± 1a 6.5 ± 0.3a 1.48 ± 0.03 
Actin–Tm − 1.4 ± 0.5b 4.6 ± 0.4 3.3 ± 0.4 1.61 ± 0.06 
Actin–Tm 3.6 ± 0.7a,c 4.5 ± 0.3c 6.6 ± 0.4a 1.46 ± 0.04 

Data are mean ± SE (n > 4) for a binding curve of eight concentrations of C0–C2 (0-20 µM) to 1 µM of actin (Fig. 1) fit to a quadratic equation. Significant difference due to PKA treatment (− versus + PKA rows).

a

P < 0.05. Significant difference due to presence of Tm (actin versus actin–Tm rows of same PKA condition).

b

P = 0.10. Units for Bmax are lifetime change (maximal Δτ) for TR-F and fraction of actin bound (C0–C2/actin) for cosedimentation.

c

P < 0.05. Trending difference in Kd due to presence of Tm. See Discussion for a detailed discussion of Kd and Bmax uses and limitations.

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