Table 2.

Summary of kinetic parameters determined by single-molecule analysis and reaction scheme

Kinetic parameters	kona	koffNPa,b	konP/konNPa,c	ktrd	kpce	Pd,f	koffPd	kcatg
	m−1 μm−1 s−1	s−1	nP/nNP	nm/s	s−1		s−1	s−1
WT	8.2 × 108 ± 3.0 × 107	1.2	0.074 ± 0.0041	52 ± 0.53	50	30	1.4 ± 0.26	2.9
F232W/F396W	9.1 × 108 ± 4.3 × 107	0.99	0.076 ± 0.0089	51 ± 0.72	49	52	0.82 ± 0.089	4.1

^a k_on, k_on^P/k_on^NP, and k_off^NP values were obtained from single-molecule fluorescence imaging analysis.

^b k_off^NP, which was the average k_off, was assumed to only contain nonproductive binding.

^c k_on^P/k_on^NP or n^P/n^NP was denoted as the productive binding ratio, where n^P and n^NP represent the number of moving and non-moving molecules for a total of 40 s observation, respectively. For the analysis, we defined moving molecules as the molecules which showed movements larger than 20 nm for 3 or more frames.

^d k_tr, processivity (P), and k_off^P were obtained by HS-AFM observation. The molecules that showed movements for 3 or more frames were analyzed.

^e k_tr was used to denote the translational velocity. k_pc was calculated by dividing the k_tr value by the step size (a product size, 1.04 nm).

^f P was estimated from the run length divided by the step size (a product size, 1.04 nm).

^g k_cat was calculated from Equation 3.