. Author manuscript; available in PMC: 2010 Apr 21.

Published in final edited form as: Biochemistry. 2009 Apr 21;48(15):3378–3386. doi: 10.1021/bi900057g

Table 2.

Kinetic Constants of Abl variants^a

	[Imidazole] (mM)	Abltide^b			ATP^c
		k_cat(s⁻¹)	K_m(µM)	k_cat/K_m(M⁻¹ s⁻¹)	k_cat(s⁻¹)	K_m(µM)	k_cat/K_m(M⁻¹ s⁻¹)
wild-type AblKD	0	12 ± 1	60 ± 20	190,000 ± 50,000	9.9 ± 0.2	45 ± 3	220,000 ± 20,000
wild-type AblKD	40	12.7 ± 0.9	38 ± 9	330,000 ± 80,000	12.5 ± 0.2	41 ± 2	300,000 ± 20,000
R367A AblKD	0	NS (> 0.04)	NS (> 500)	35 ± 9
R367A AblKD	40	8.9 ± 0.3	130 ± 20	68,000 ± 8,000	5.6 ± 0.1	47 ± 5	120,000 ± 10,000
A365R/R367A AblKD	0	22 ± 1	280 ± 30	80,000 ± 10,000	13.2 ± 0.4	114 ± 9	115,000 ± 9,000

Reactions were performed at 30 ° in 100 mM Tris, pH 7.4 and 10 mM MgCl₂. NS: No saturation. If no value is given, then the constants were not measured.

For wild-type AblKD and R367A AblKD reactions without imidazole: [γ−³²P]ATP = 0.15 mCi/mL and ATP = 0.25 mM ATP. For wild-type AblKD with imidazole: [γ−³²P]ATP = 0.15 mCi/mL and ATP = 1 mM ATP. Otherwise, reaction mixtures contained 1 mM ATP, 0.1 U/µL pyruvate kinase, 0.05 U/µL lactate dehydrogenase, 1 mM phosphoenolpyruvate, 150 µM NADH, and 0.5 mM Na₃VO₄

Reaction mixtures included 0.1 U/µL pyruvate kinase, 0.05 U/µL lactate dehydrogenase, 1 mM phosphoenolpyruvate, 150 µM NADH, and 0.5 mM Na₃VO₄. For wild-type AblKD, [Abltide] = 190 µM; for R367A AblKD, [Abltide] = 440 µM; for A365R/R367A AblKD, [Abltide] = 840 µM.