Table 1. Summary of Trp fluorescence parameters measured for Ail-WT and its Trp mutants in various DPRs.

	Average lifetime (<τ>, ns)b			Anisotropy (r)b			Bimolecular quenching constant (kq, × 109 M−1 s−1)
DPRa	WT	W42F	W149F	WT	W42F	W149F	WT	W42F	W149F
700:1	3.62 ± 0.2	2.43 ± 0.2	3.17 ± 0.1	0.143	0.122	0.152	1.40	1.54	1.85
700:1 (D)	1.96 ± 0.0	1.68 ± 0.1	1.77 ± 0.1	0.068	0.070	0.062	-	-	-
1750:1	3.96 ± 0.1	2.93 ± 0.2	3.72 ± 0.3	0.140	0.123	0.159	1.33	1.36	1.77
1750:1(D)	2.10 ± 0.1	1.75 ± 0.1	1.87 ± 0.1	0.080	0.068	0.072	-	-	-
3500:1	3.97 ± 0.1	2.89 ± 0.1	4.34 ± 0.1	0.141	0.120	0.154	1.31	1.35	1.56
3500:1 (D)	2.18 ± 0.1	1.81 ± 0.0	1.86 ± 0.0	0.106	0.080	0.069	-	-	-
0:1 (U)	1.75 ± 0.1	1.66 ± 0.1	1.61 ± 0.1	0.050	0.046	0.047	4.18	4.31	4.78
0:1 (A)	0.32 ± 0.0c	0.14 ± 0.0c	0.20 ± 0.0c	0.123	0.143	0.142	-c	-c	-c

^aHere (D) represents the refolded protein denatured by the addition of 6.4 M GdnHCl, and therefore retains the respective LDAO amounts. (U) is unfolded protein in 8.0 M GdnHCl and contains no LDAO. (A) is aggregated protein sample prepared in Tris-HCl pH 8.5 and contains 0.16 M GdnHCl.

^bAll samples were incubated for 48 h at 25°C before obtaining measurements. <τ> = Στ = α_iτ_i

^cLifetimes could not be accurately determined due to interference from scattering. Hence, the k_q values were also not calculated.