Protein Engineering, Vol. 12, No. 3, 189-202,
March 1999
© 1999 Oxford University Press
Solution structure and dynamics of a serpin reactive site loop using interleukin 1ß as a presentation scaffold
Department of Chemistry, Brandeis University, Waltham, MA 02454-9110 and 4 Department of Chemistry, Wellesley College, Wellesley, MA 02181-8284, USA
Human interleukin-1ß (IL1ß) was used as a presentation scaffold for the characterization of the reactive site loop (RSL) of the serpin 
1-antitrypsin (A1AT), the physiological inhibitor of leukocyte elastase. A chimeric protein was generated by replacement of residues 50–53 of IL1ß, corresponding to an exposed reverse turn in IL1ß, with the 10-residue P5-P5' sequence EAIPMSIPPE from A1AT. The chimera (antitrypsin-interleukin, AT-IL) inhibits elastase specifically and also binds the IL1ß receptor. Multinuclear NMR characterization of AT-IL established that, with the exception of the inserted sequence, the structure of the IL1ß scaffold is preserved in the chimera. The structure of the inserted RSL was analyzed relative to that of the isolated 10-residue RSL peptide, which was shown to be essentially disordered in solution. The chimeric RSL was also found to be solvent exposed and conformationally mobile in comparison with the IL1ß scaffold, and there was no evidence of persisting interactions with the scaffold outside of the N- and C-terminal linkages. However, AT-IL exhibits sigificant differences in chemical shift and NOE patterns relative to the isolated RSL that are consistent with local features of non-random structure. The proximity of these features to the P1-P1' residues suggests that they may be responsible for the inhibitory activity of the chimera.
Keywords: serpin/1-antitrypsin/interleukin-1ß/protease inhibitors
1 Present address: Department of Chemistry, PRAECIS Pharmaceuticals, Inc., 1 Hampshire Street, Cambridge, MA 02139, USA
2 Present address: Department of Chemistry and Biochemistry, Northwestern University, Evanston IL 60208-3113, USA
3 To whom correspondence should be addressed
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