In their 2010 study, Consequently, an increase in Λ corresponds to a lengthening of the 𝜆i distances and decreased hydrogen bonding in the β-hairpin. The other parameter used to define phase space was the Glu8 Ψ angle, which was estimated to be around -80° for right-handed structures and 90° for left ones. Using the same parameters, we evaluated our simulations and found similar dominant Ψ angles of -24°/-33° and 155°/145° for structures with/without the isopeptide bond intact, suggesting even after the bond is broken the pre-lasso retains right-handed structures (Figure ). However, it was not clear if the Glu8 Ψ angle could delineate handedness. This motivated our development of the lasso handedness descriptor, 𝜅, which definitively distinguishes right-and left-handed structures (Figure ). This descriptor confirmed that no left-handed configurations were obtained in any of our ten replicas. In the previous work, lefthanded structures remained stable for 30 ns with Λ = 6.2 ± 0.3 Å and Glu8 Ψ = 90±20°. Filtering our ensembles through Λ values, we found a considerable diversity of pre-tadpoles and unfolded structures, but no left-handed pre-lassos.