Table 2 Four different types of fluorescence measurements were used to measure NADH binding affinity for rmGAPDH and byGAPDH.

From: Substrate Channeling via a Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

 

rmGAPDH-NADH KD fluorescence measurements

byGAPDH-NADH KD fluorescence measurements

Measurement type

KD in micro Molar + error

KD in micto molar + error

Protein quenchinga

0.84 ± 0.09

8.7 ± 0.6

NADH quenchingb

0.86 ± 0.07

8.5 ± −0.8

FRET protein-NADHc

0.77 ± 0.5

8.0 ± 0.7

Anisotropy NADHd

0.73 ± 0.06

7.6 ± 0.5

Average:

0.8 ± 0.06

8.2 ± 0.5

  1. The KD constants are shown in terms of NADH binding sites on each tetramer:
  2. aexcitation 295 nm emission 328 nm; bexcitation 328 nm emission 457 nm; cexcitation 295 nm emission 457 nm; dexcitation 330 nm emission 457 nm.
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