BiKK structure

Supplementary material

NMR structure note:Solution structure of anovel C2-symmetrical bifunctional bicyclic inhibitor based

on SFTI-1

Agnès M. Jaulenta, Arnd B.E. Brauera,b, Stephen J. MatthewscRobin J. Leatherbarrow1*

aDepartment of Chemistry and cDepartment of Biological Sciences, Imperial College London, South Kensington Campus, LondonSW7 2AZ, U.K.

bFreie Universität Berlin, Institut für Chemie – Biochemie, Otto-Hahn-Bau, Thielallee 63, D-14195 Berlin (Dahlem), Germany

*Email:

Table 1.Assignment table for the chemical shifts in ppm of the CTKSIPP’I’repeat unit of BiKK, including coupling constants and derived dihedral restraints.

HN /  /  /  /  / other H / 3JHNH
in Hz / 3JHH
in Hz / 
in o / 1
in o
Cys / 8.92 / 5.89 / 2.94 S
3.04 R / 10.6 / 2.8
12.4 / -120±20 / -60±30
Thr / 8.71 / 4.43 / 4.61 / 1.43 / 7.0 / 2.9 / -120±60 / +60±30
Lys / 8.51 / 4.50 / 2.02
2.06 / 1.51
1.43 / 1.70 / 3.00
HN7.49 / 8.3 / 7 / -120±45
Ser / 7.29 / 4.39 / 3.79
3.90 / 7.8 / 3.7
3.3 / -120±45 / +60±30
Ile / 8.21 / 4.31 / 1.80 / γ1 1.45
1.07
γ2 0.84 / 0.84 / 10.0 / 11 / -120±20 / -60±30
Pro / - / 5.10 / 2.46 S
2.04 R / 1.85 S
1.96 R / 3.63 S
3.54 R / - / 8.9
3.0 / -70±20 / +26±15
Pro′ / - / 4.22 / 2.45 S
1.87 R / 2.04 S
2.11 R / 3.83 S
3.72 R / - / 9.6
4.5 / -70±20
Ile′ / 7.87 / 4.34 / 1.93 / γ1 1.46
1.16
γ2 0.87 / 0.81 / 10.0 / 11 / -120±20 / -60±30

Table 2.Statistics of the family of 20lowest-energy structures calculated for the bifunctional trypsin inhibitor

Distance restraints for each C-T-K-S-I-P-P'-I'- segment
Intraresidue / 34
i + 1 / 19
i + 2 / 3
i + 3 / 1
i + 4 / 1
i + 5 / 1
i + 6 / 5
i + 7 / 1
Dihedral restraints for each C-T-K-S-I-P-P'-I'- segment
 / 8
 / 8
1 / 6
Hydrogen bond restraints / 0
Ramachandran analysis
Residues in favoured regions / 83.2 %
Residues in additional allowed regions / 16.8 %
Residues in generously allowed regions / 0 %
Residues in disallowed regions / 0 %
Pairwise root mean square deviations (Å)
All residues over backbone atoms / 0.24  0.08 (0.00…0.47)
All residues over heavy atoms / 1.05  0.25 (0.00…1.63)
Backbone atoms of the two C-T-K-S-I-P-P'-I'- segments / 0.18  0.07 (0.00…0.33)
Heavy atoms of the two C-T-K-S-I-P-P'-I'- segments / 0.98  0.30 (0.00…1.71)
Pairwise root mean square deviations from SFTI-1 over C-T-K-S-I-P-P'-I'-C loop (Å)
Crystal structure in complex with trypsin (1SFI)
Backbone atoms / 0.45  0.03 (0.42…0.51)
Heavy atoms / 1.16  0.25 (0.77…1.54)
NMR solution structure (1JBL)
Backbone atoms / 0.31  0.15 (0.06…0.61)
Heavy atoms / 0.98  0.25 (0.37…1.60)
11-mer BBI mimic (1GM2)
Backbone atoms / 0.4  0.13 (0.08…0.70)

- 1 -