Table 1.

Measured and Predicted Hydrodynamic Parameters for NSF-WT or Mutants in ADP- and AMP-PNP-Bound States. (Ni²⁺-NTA)₂-Cy3-His₆-NSF-Myc wild-type or mutants were in Buffer A containing 0.5 mM ADP or AMP-PNP. Sedimentation velocity was performed at 15,000 rpm and 10° C. Radial intensity scan data were collected, converted to 2 pseudoabsorbance data and fit to the c(s) model by SEDFIT program. The peak s-values have been corrected to reflect water density and viscosity at 20° C (s_20,W). The experiments were repeated thrice, all the values in the table were averaged and indicated with the subscript meas. The models in Fig. 2 and the crystal structure of NSF-D2 (PDB entry 1D2 N) were evaluated using the Solution Modeler (SOMO) routine (Fig. 3; Brookes et al., 2010a,b) and hydrodynamic parameters predicted from that analysis are indicated with the subscript pred. Values predicted for the most extended intermediate obtained by morphing between ADP and ATP forms (Movie 3, frame 00:00:04) are indicated with the subscript pred, int. % Hexamer, the percentage of hexamer in solution; s_20,W, sedimentation coefficient; ΔAMP-PNP-ADP, s_20,W values of ADP state subtracted from that of AMP-PNP bound state; f/f₀, frictional ratio; MW, molecular weight.

	% Hexamer		s20,W		Δ	f/f0		MW (kDa)
	ADP	AMP-PNP	ADP	AMP-PNP	AMP-PNP-ADP	ADP	AMP-PNP	ADP	AMP-PNP
NSFmeas	62.01 ± 5.74	46.99 ± 3.70	11.43 ± 1.00	12.11 ± 1.20	0.68	1.54 ± 0.01	1.46 ± 0.02	525 ± 92	511 ± 95
NSFpred	–	–	16.37	16.91	0.54	1.33	1.28	–	–
NSFpre,int	–	–	–	14.22	–	–	1.37	–	–
D1D2meas	43.99 ± 2.93	14.10 ± 0.31	10.13 ± 1.09	8.13 ± 0.92	−2.00	1.37 ± 0.001	1.66 ± 0.03	332 ± 77	290 ± 76
D1D2pred	–	–	13.89	–	–	1.27	–	–	–
D2meas	28.84 ± 1.92	10.30 ± 0.63	6.66 ± 0.43	6.59 ± 0.39	−0.07	1.18 ± 0.003	1.19 ± 0.02	131 ± 18	151 ± 19
D2pred	–	–	–	7.10	–	–	1.31	–	–
L441Ameas	34.44 ± 1.06	34.19 ± 2.78	9.94 ± 1.58	9.93 ± 1.34	30.01	1.77 ± 0.07	1.78 ± 0.07	474 ± 141	530 ± 144
R67A meas	55.59 ± 12.48	38.71 ± 4.21	11.10 ± 0.96	11.85 ± 1.12	0.75	1.59 ± 0.02	1.49 ± 0.004	543 ± 86	506 ± 87