Table 1 Equilibrium and kinetic constants of tubulin complexes with DARPin D1 and its variants.

From: Destabilizing an interacting motif strengthens the association of a designed ankyrin repeat protein with tubulin

DARPins

D1

A-C2

TM-3

C-cap-truncated TM-3

Fluorescence data

   

(Digested TM-3)

 KD (titration) (nM)

127 ± 12

0.82 ± 0.18

0.90 ± 0.19

1.03 ± 0.28

 koff (s−1)

1.49 ± 0.07

0.0071 ± 0.0002

0.0049 ± 0.0002

0.0046 ± 0.0001

 kon (M−1s−1)

1.30 × 107

1.60 × 107

1.82 × 107

2.67 × 107

 koff/kon (nM)

115 ± 6

0.444 ± 0.013

0.269 ± 0.013

0.172 ± 0.004

SPR data

   

(TM-3 1–149)

 KD (titration) (nM)

300 ± 30

n.d.

n.d.

n.d.

 koff (s−1)

0.22 ± 0.01

0.0041 ± 0.0001

0.0025 ± 0.0002

0.0027 ± 0.0001

 kon (M−1s−1)

8 ± 1.5 × 105

1.2 ± 0.2 × 106

1.9 ± 0.2 × 106

1.7 ± 0.1 × 106

 koff/kon (nM)

275 ± 75

3.4 ± 0.6

1.3 ± 0.2

1.6 ± 0.2

  1. n.d.: not determined.
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