Table 2.

Kinetic parameters for wild-type and mutant Ty3 RT variants

	Enzyme	KmdTTP (μM)	Kcat (s−1)	Kcat/Km (μM−1 S−1)
poly(rA):oligo(dT)15	WT	14	2.4	0.18
	Asp151→Glu	690	2.1 × 10−5	0.3 × 10−7
	Asp213→Glu	510	0.15	0.29 × 10−3
	Asp214→Asn	34	0.02	0.59 × 10−3
	Asp214→Glu	78	0.031	0.41 × 10−3
	Asp214→Gln	26	0.0016	0.62 × 10−4
	Enzyme	KmdGTP (μM)	Kcat (s−1)	Kcat/Km (μM−1 S−1)
poly(rC):oligo(dG)18	WT	42	6.2	0.15
	Asp151→Glu	ND	ND	ND
	Asp213→Glu	260	0.67	0.26 × 10−2
	Asp214→Asn	42	0.12	0.28 × 10−2
	Asp214→Glu	64	0.19	0.3 × 10−2
	Asp214→Gln	ND	ND	ND
poly(dC):oligo(dG)18
	WT	19	24	1.3
	Asp151→Glu	ND	ND	ND
	Asp213→Glu	140	0.88	0.64 × 10−2
	Asp214→Asn	4	0.044	0.012
	Asp214→Glu	14	0.12	0.86 × 10−2
	Asp214→Gln	ND	ND	ND

The steady-state parameters were derived using the template/primer indicated and corresponding dNTP substrate as described in Materials and Methods. ND, not determined.