TABLE 1.

Substrate binding of wild type and mutant VCB complexes The K_d and k_off values for the interaction with the HIF-1α-derived Hyp-MIANS peptide (15) were determined by fitting the data shown in supplemental Fig. S2 as described (15).

	Kd	koff	kona
	nm	s–1	m–1 s–1
Wild type	5.7 ± 0.5	0.018 ± 0.00012	3.2 × 106
P81S	15.8 ± 1.8	0.023 ± 0.00016	1.4 × 106
L188V	18.1 ± 1.6	0.018 ± 0.00019	0.99 × 106
P81S/L188V	21.6 ± 3.6	0.021 ± 0.00059	0.97 × 106
V84L	NDb	0.014 ± 0.00018	NDb

^aThe k_on values were calculated using the equation k_on = k_off/K_d

^bND, not determined. K_d could not be determined because limiting yields of recombinant pVHL^V84L-containing VCB complex precluded the preparation of meaningful titration series