. Author manuscript; available in PMC: 2020 Oct 15.

Published in final edited form as: Biochemistry. 2019 Oct 7;58(41):4260–4268. doi: 10.1021/acs.biochem.9b00732

Table 2.

Kinetic parameters of T104X mutants of 17X-PTDH^a

Variant	k_cat^b (s⁻¹)	K_m,NAD^b (mM)	K_m,PT^b (mM)	K_iA,NAD^b (mM)	k_cat/K_m,PT^b (x 10⁴ M⁻¹s⁻¹)	^Dk_cat^c	^D(k_cat/K_m,PT)^c
17X	2.53 (0.07)	0.036 (0.005)	0.061 (0.006)	0.47 (0.06)	4.1 (0.4)	2.27 (0.05)	2.3 (0.1)
T104S	1.31 (0.02)	0.042 (0.007)	0.13 (0.01)	2.1 (0.3)	1.0 (0.1)	2.27 (0.01)	2.2 (0.1)
T104A	1.02 (0.03)	0.023 (0.002)	0.27 (0.02)	0.31 (0.03)	0.38 (0.03)	2.27 (0.07)	2.1 (0.2)
T104G	0.31 (0.01)	0.035 (0.003)	0.31 (0.02)	0.85 (0.07)	0.10 (0.01)	2.19 (0.02)	2.1 (0.1)

All parameters were obtained at 25 °C and pH 7.25 in 100 mM MOPS.

Values determined using variable concentrations of ¹H-PT and NAD⁺.

Values determined using saturating concentrations of NAD⁺ and varied concentrations of ¹H- and ²H-PT.