2 research outputs found
Human Leukocyte Elastase Inhibitors: Designed Variants of Human Pancreatic Secretory Trypsin Inhibitor (hPSTI)
DESIGN OF EFFICIENT HUMAN LEUKOCYTE ELASTASE INHIBITORS; VARIANTS OF HUMAN PANCREATIC SECRETORY TRYPSIN INHIBITOR (hPSTI)
A set of hPSTI variants were constructed by total gene synthesis
or site-specific mutagenesis, with the aim of producing human
leukocyte elastase(HLE)-specific inhibitors and obtaining a better
insight into the parameters effecting inhibitor/protease interaction.
In the initial planning the structure of related Kazal-type
inhibitors were taken into account. For subsequent protein design,
models were made of HLE based on the structure of porcine pancreatic
elastase, and of hPSTI based on the stucture of porcine PSTI. Models
of the hPSTI/HLE and hPSTI/chymotrypsin complexes were generated by
CAPD "docking". The modelled complexes could be used to rationalise a
posteriori the inhibitory properties of the hPSTI variants, and to
postulate the structure of better elastase inhibitors which were duly
generated. Excellent specific inhibitors (Ky=1.5x107 4" M for SHEE)
were obtained and the contribution of individual amino acid residues
Or exchanges to the binding constant were estimated