Abstract
A novel bifunctional bicyclic inhibitor has been created that combines features both from the Bowman–Birk inhibitor (BBI) proteins, which have two distinct inhibitory sites, and from sunflower trypsin inhibitor-1 (SFTI-1), which has a compact bicyclic structure. The inhibitor was designed by fusing together a pair of reactive loops based on a sequence derived from SFTI-1 to create a backbone-cyclized disulfide-bridged 16-mer peptide. This peptide has two symmetrically spaced trypsin binding sites. Its synthesis and biological activity have been reported in a previous communication [Jaulent and Leatherbarrow, 2004, PEDS 17, 681]. In the present study we have examined the three-dimensional structure of the molecule. We find that the new inhibitor, which has a symmetrical 8-mer half-cystine CTKSIPP′I′ motif repeated through a C2 symmetry axis also shows a complete symmetry in its three-dimensional structure. Each of the two loops adopts the expected canonical conformation common to all BBIs as well as SFTI-1. We also find that the inhibitor displays a strong and unique structural identity, with a notable lack of minor conformational isomers that characterise most reactive site loop mimics examined to date as well as SFTI-1. This suggests that the presence of the additional cyclic loop acts to restrict conformational mobility and that the deliberate introduction of cyclic symmetry may offer a general route to locking the conformation of β-hairpin structures.
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Abbreviations
- 1D:
-
One-dimensional
- 2D:
-
Two-dimensional
- BBI:
-
Bowman–Birk inhibitor
- BiKK:
-
cyclo[(CTKSIPPI)2] with disulfide bridge
- NOESY:
-
Nuclear Overhauser effect spectroscopy
- RMSD:
-
Root mean square deviation
- SFTI-1:
-
sunflower trypsin inhibitor-1, cyclo(CTKSIPPICFPDGR) with disulfide bridge
- TOCSY:
-
Total correlation spectroscopy
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Jaulent, A.M., Brauer, A.B.E., Matthews, S.J. et al. Solution Structure of a Novel C2-Symmetrical Bifunctional Bicyclic Inhibitor Based on SFTI-1. J Biomol NMR 33, 57–62 (2005). https://doi.org/10.1007/s10858-005-1210-9
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DOI: https://doi.org/10.1007/s10858-005-1210-9