Title: Nucleotide-dependent conformational changes in a protease-associated ATPase HslU

Authors: J. Wang, J.J. Song, I.S. Seong, M.C. Franklin, S. Kamtekar, S.H. Eom, and C.H. Chung.

Supplementary Movies.

All movies were composed of images made using SPOCK (J.A. Christopher, 1998. SPOCK: The structural properties observation and calculation kit. College Station, Texas, The Center for Macromolecular Design, Texas, A&M University). With the exception of Movie 2f, which was made of 22 images forward and backward, all movies were made of 50 images in each direction. For nucleotide binding-induced conformational changes, the coordinates of 1G4A were used for the 6 ADP-bound state, the coordinates of 1DO2 were used for the 3 ADP-bound state, and all intermediate coordinates were linearly iStatenterpolated between these two sets of coordinates without energy minimization. Missing parts in domain I in any one of the two structures were excluded in calculations. For ATP hydrolysis-induced conformational changes, the coordinates of 1G4A were used for the 6 ADP-bound state, and the coordinates of 1G3I were used for the 6 ATP-bound state. Before linear interpolation of intermediate coordinates, mutations were made in the ATP-bound state using the first rotamer in the side chain rotamer library so that the two states would have identical sequences. Additionally, a part of domain I that was present in the ADP-bound state but absent in the ATP-bound state was added back to the ATP-bound state using local least squares superposition of remaining parts of this domain. Unlike the other movies, the interpolation for Movie 2f was carried out using Morph (M. Gerstein, W.G. Krebs, 1998, A database of macromolecular motions. Nuc. Acid. Res. 26, 4280-4290) with energy minimization. In the Morph interpolation, image 1 and image 22 were the observed ATP (starting)- and ADP (ending)-bound states, respectively; image 2 was an energy-minimized state that was 1/21th interpolated between images 1 and 22; image 3 was an energy-minimized state that was 1/20th interpolated between images 2 and 22, and so on. Due to the requirement for a single chain in the current version of this program, the ends of missing domain I were joined together with energy minimized.

Movie 1. Nucleotide binding-induced conformational changes in HslU

Movie 1a. This movie is complementary to Figure 3 and shows nucleotide binding-induced conformational changes in hexameric HslU between the 6 ADP and 3 ADP bound states viewed from the apical sufrace.

Movie 1b. This movie is complementary to Figure 3 and shows nucleotide binding-induced conformational changes in hexameric HslU between the 6 ADP and 3 ADP bound states viewed from the domain I direction.

Movie 1c. This movie is complementary to Figure 3 and shows nucleotide binding-induced conformational changes in hexameric HslU between the 6 ADP and 3 ADP bound states viewed from side.

Movie 1d. This movie is complementary to Figure 3 and shows nucleotide binding-induced conformational changes in hexameric HslU between the 6 ADP and 3 ADP bound states viewed from the apical surface. This movie focuses on two adjacent subunits within hexameric HslU and shows a "riding" motion.

Movie 1e. This movie is complementary to Figure 3 and shows nucleotide binding-induced conformational changes in hexameric HslU between the 6 ADP and 3 ADP bound states viewed from side. This movie focuses on two adjacent subunits within hexameric HslU. Motions in this movie much resembles a running dog (Gibbs, Professor D. M. Engleman's big dog at Yale University).

Movie 2. ATP hydrolysis induced conformation changes in HslVU and HslU

Movie 2a. This movie is complementary to Figure 5 and shows ATP hydrolysis -indueced conformational changes in the HslVU complex viewed from side.

Movie 2b. This movie is complementary to Figure 5 and shows ATP hydrolysis -indueced conformational changes in the HslVU complex viewed from a tilt angle.

Movie 2c. This movie is complementary to Figure 4 and shows ATP hydrolysis -indueced conformational changes in HslU viewed from side.

Movie 2d. This movie is complementary to Figure 4 and shows ATP hydrolysis -indueced conformational changes in HslU viewed from the apical surface.

Movie 2e. This movie is complementary to Figure 4 and shows ATP hydrolysis -indueced conformational changes in HslU viewed from the domain I direction.

Movie 2f. This movie is complementary to Figure 4 and shows ATP hydrolysis -indueced conformational changes in single subunit of HslU viewed from inside.