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. Author manuscript; available in PMC: 2019 Jan 26.
Published in final edited form as: Biochemistry. 2018 Nov 30;57(50):6888–6896. doi: 10.1021/acs.biochem.8b01134

Table 2.

Kinetic Rate Constant Estimates determined by Surface Plasmon Resonancea

Protein kon (M−1s−1) SD koff (s−1) SD Kd (nM) errorb Ki (nM)c Fold differencee
EapH1 WT 2.23×105 5.0×103 6.03×10−5 3.1×10−6 0.271 0.015 0.021 12.9
EapH1 WTd 4.85×105 2.06×105 7.10×10−5 5.6×10−6 0.146 0.063 0.021 7.0
R89M 2.85×104 5.13×103 0.0737 0.0115 2586 615 321 8.1
R89K 8.72×104 7.91×103 0.0165 0.0020 189 29 26.8 7.0
R89Q 2.32×104 2.32×103 0.1397 0.0271 6028 1316 944 6.4
R89M/E94Q 5.27×104 2.99×103 0.0876 0.0084 1662 185 236 7.0
R89M/K95M 2.49×104 4.61×103 0.0904 0.0166 3632 945 437 8.3
R89M/E94Q/K95M 2.08×104 1.14×103 0.0855 0.0124 4108 637 406 10.1
a

Microscopic rate constant estimates are the average of either four individual sensograms (WT) or six individual sensograms (mutants).

b

The dissociation constant error was propagated from the on- and off-rate standard deviations.

c

The inhibition constants determined by progress curve analysis (WT) or 5×5 assay (R89M mutants, Fig. 4).

d

Microscopic rate constant estimates determined by single-cycle binding kinetics.

e

The fold difference depicts the difference between the dissociation constant determined by SPR and the inhibition constant determined by kinetic analysis.