We find that moving along the first tIC generated by the concatenated trajectory corresponds to transitioning between the closed, intermediate and open crystal structures (Figure ). To better understand how regions of the TICA space correspond to different F-helix conformations, we project the three native crystal structures (4w52, 4w57 and 4w59) onto our 2D TICA surface. For each native crystal structure, we calculate the relevant C α -C α and C β -C β pairwise distances and use those features to calculate coordinates in TICA space. We find that the open crystal structure (4w59) is roughly centered in the larger half-moon region on the left of the TICA space, the intermediate crystal structure (4w57) is right of the bridge region between wells and the closed crystal structure (4w52) is centered in the ovalshaped well on the right (Figure ). Consistent with what was reported in experiment, we find from our TICA projections that the intermediate crystal structure falls between the closed and open crystal structures. Based on the position of each crystal structure in the 2D TICA surface, we determine that the differences among discrete states is predominately captured by variation in the first independent component (Figure ). Time-lagged independent component analysis provides an alternative to RMSD for defining discrete states. We find that our native system trajectories generally occupy distinct regions of the TICA space. Using the tICs we previously define, we map each native system trajectory onto the 2D TICA surface. We observe that all frames of simulations of the 4w52-benzene system map to the right side of the oval-shaped well and center around the 4w52 crystal structure (Figure ). Simulations of the 4w57-butylbenzene system sample all regions of the oval-shaped well in addition to the bridge region linking the two wells together (Figure ). Lastly, simulations of the 4w59-hexylbenzene system exclusively sample the half-moon region on the left side of the TICA plot (Figure ). By contrasting the different regions of TICA space these systems occupy, we posit that the experimentally observed closed conformation is represented by the right oval-shaped well, the experimentally observed open conformation is the left half-moon well and the experimentally observed intermediate conformation is the bridge region between them in the 2D TICA space (Figure ).