| cMyBP-C complex with 1 µM F-actin . | Change in IAEDANS lifetime (%) . | ||
|---|---|---|---|
| Construct . | IAEDANS . | −PKA . | +PKA . |
| C0–C2 WT (2.5 µM) | Actin | 2.61 ± 0.14 | 0.88 ± 0.08 |
| C0–C2 WT (1.25 µM) | Actin | 1.92 ± 0.17 | 0.68 ± 0.08 |
| C0–C2 WT (0.625 µM) | Actin | 0.83 ± 0.1 | |
| C0–C2 WT (1.25 µM) | Actin–Tm | 2.70 ± 0.07 | 0.71 ± 0.06 |
| C0–C2 WT (0.65 µM) | Actin–Tm | 1.70 ± 0.12 | |
| C0–C2 WT (0.0313 µM) | Actin–Tm | 0.56 ± 0.06 | |
| C0–C1 (Δm-C2; 2.5 µM) | Actin | 0.17 ± 0.06 | |
| C0–C2 L352P (2.5 µM) | Actin | 3.62 ± 0.19 | |
| C0–C2 E334K (2.5 µM) | Actin | 2.79 ± 0.11 | |
| C0–C2 R282W (2.5 µM) | Actin | 2.20 ± 0.24 | 0.86 ± 0.12 |
| C0–C2 R282W (1.25 µM) | Actin | 1.24 ± 0.12 | 0.53 ± 0.08 |
| C0–C2 R215E/K218E (1.25 µM) | Actin | 0.77 ± 0.06 | |
| C0–C2 R215E/K218E (1.25 µM) | Actin–Tm | 0.48 ± 0.05 | |
| C0–C2 A216R/S217K (1.25 µM) | Actin | 2.63 ± 0.10 | |
| C0–C2 A216R/S217K (0.625 µM) | Actin | 2.16 ± 0.13 | |
| C0–C2 A216R/S217K (1.25 µM) | Actin–Tm | 2.98 ± 0.08 | |
| C0–C2 A216R/S217K (0.625 µM) | Actin–Tm | 2.68 ± 0.05 | |
| C0–C2 A216R/S217K (0.313 µM) | Actin–Tm | 2.08 ± 0.12 | |
| cMyBP-C complex with 1 µM F-actin . | Change in IAEDANS lifetime (%) . | ||
|---|---|---|---|
| Construct . | IAEDANS . | −PKA . | +PKA . |
| C0–C2 WT (2.5 µM) | Actin | 2.61 ± 0.14 | 0.88 ± 0.08 |
| C0–C2 WT (1.25 µM) | Actin | 1.92 ± 0.17 | 0.68 ± 0.08 |
| C0–C2 WT (0.625 µM) | Actin | 0.83 ± 0.1 | |
| C0–C2 WT (1.25 µM) | Actin–Tm | 2.70 ± 0.07 | 0.71 ± 0.06 |
| C0–C2 WT (0.65 µM) | Actin–Tm | 1.70 ± 0.12 | |
| C0–C2 WT (0.0313 µM) | Actin–Tm | 0.56 ± 0.06 | |
| C0–C1 (Δm-C2; 2.5 µM) | Actin | 0.17 ± 0.06 | |
| C0–C2 L352P (2.5 µM) | Actin | 3.62 ± 0.19 | |
| C0–C2 E334K (2.5 µM) | Actin | 2.79 ± 0.11 | |
| C0–C2 R282W (2.5 µM) | Actin | 2.20 ± 0.24 | 0.86 ± 0.12 |
| C0–C2 R282W (1.25 µM) | Actin | 1.24 ± 0.12 | 0.53 ± 0.08 |
| C0–C2 R215E/K218E (1.25 µM) | Actin | 0.77 ± 0.06 | |
| C0–C2 R215E/K218E (1.25 µM) | Actin–Tm | 0.48 ± 0.05 | |
| C0–C2 A216R/S217K (1.25 µM) | Actin | 2.63 ± 0.10 | |
| C0–C2 A216R/S217K (0.625 µM) | Actin | 2.16 ± 0.13 | |
| C0–C2 A216R/S217K (1.25 µM) | Actin–Tm | 2.98 ± 0.08 | |
| C0–C2 A216R/S217K (0.625 µM) | Actin–Tm | 2.68 ± 0.05 | |
| C0–C2 A216R/S217K (0.313 µM) | Actin–Tm | 2.08 ± 0.12 | |
Substoichiometric binding levels of cMyBP-C C0–C2 and C0–C1 on 1 µM IAEDANS-labeled actin and actin–Tm. Data are provided as mean ± SE (n > 4). P values comparing different samples are given in Results.