Table 1. Structural statistics of PRE-based structural ensembles.
| restraints for β-interface | restraints for α-interface at S1’ , V-B loops | |
|
| ||
| Restraints$ to 5-Doxyl position | ||
|
| ||
| HN 8a aa *8 DMPC lipids *2 acyl chains = 128 | HN 9e aa *8 DMPC lipids *2 acyl chains = 144 | |
|
| ||
| HCH3 8b aa *8 DMPC lipids *2 acyl chains = 128 | HCH3 7f aa *8 DMPC lipids *2 acyl chains =112 | |
|
| ||
| Restraints$ to 14-Doxyl position | ||
|
| ||
| HN 4c aa *8 DMPC lipids *2 acyl chains = 64 | HN 6g aa *8 DMPC lipids *2 acyl chains = 96 | |
|
| ||
| Restraints$ to fluor-tagged position | ||
|
| ||
| CA 2d aa *8 DMPC lipids *2 acyl chains = 32 | CA 2h aa *8 DMPC lipids *2 acyl chains = 32 | |
|
| ||
| Total PRE-based distance restraints = 352 | Total PRE-based distance restraints = 384 | |
|
| ||
| aR117, N119, T154, A167, H168, D170, G190, I191 | eH206, S207, G208, K233, K241, Y242, D244, T247, D254 | |
| bT115, T154, M156, L160, V162, A167, A173, I191 | fT205, V235, V243(2), I245, T247, L250 | |
| cV162, H168, F171, I191 | gH206, G208, N211, K241, D244, T247 | |
| dG155C, G190C | hS207C, A252C | |
|
| ||
| 56 φ + 57 ψ dihedral angle restraints§ = 113 | 56 φ + 57 ψ dihedral angle restraints§ = 113 | |
| Total NMR solution structural restraints = 465 | Total NMR solution structural restraints = 497 | |
|
| ||
| Distances to metals¶:17 to Zn++ + 19 to Ca++ = 36 | Distances to metals¶:: 17 to Zn++ + 19 to Ca++ = 36 | |
|
| ||
|
RMS deviations£
|
||
| β-interface | α-interface | |
|
| ||
| Average pairwise r.m.s deviation (Å) | ||
| All Backbone atoms | 0.79 ± 0.08 | 0.75 ± 0.06 |
| All Heavy atoms | 1.25 ± 0.09 | 1.18 ± 0.07 |
| Backbone to MMP-12 NMR structure (2POJ) (Å) | 1.85 ± 0.13 | 1.71 ± 0.09 |
| RMSD from PRE-based distance restraints (Å) | 0.96 ± 0.19 | 0.93 ± 0.13 |
| Q-factor for PRE-based distance restraints | 0.06 ± 0.01 | 0.06 ± 0.01 |
| Deviations from idealized bond lengths (Å) | 0.011 ± 0.001 | 0.01 ± 0.001 |
| Deviations from idealized bond angles (°) | 3.61 ± 0.05 | 3.57 ± 0.06 |
|
|
||
| Statistics of overall structural quality |
Mean Score
|
|
| Procheck G-factor (all dihedrals) | −0.59 | −0.64 |
| Molprobity ¢ | 1.49 | 1.59 |
| Residues in most favored regions of Ramachandran plot€ | 89.8% | 92.7% |
| Residues in additional allowed regions | 8.8% | 5.2 % |
$
ambiguous with respect to location of spin-labeled phospholipid; each restraint is satisfied by proximity to any of 8 phospholipids in either acyl chain at the depth corresponding to the position of the label
§
Dihedral restraints of Bhaskaran et al. 23
¶
Based on distances measured by Lang et al. 37
£
RMSD calculated from the ensemble of 14 lowest energy structures.
¢
Molprobity score combines the clash score, rotamers, and Ramachandran evaluation into a single score.
€
from Procheck-NMR