The crystal structure of a doubleheaded
a-chymotrypsin inhibitor, WCI, from winged
bean seeds has now been refined at 2.3A°
resolution
to an R-factor of 18.7% for 9,897 reflections. The
crystals belong to the hexagonal space group P6122
with cell parameters a 5 b 5 61.8A°
and c 5 212.8A°
.
The final model has a good stereochemistry and a
root mean square deviation of 0.011A°
and 1.14° from
ideality for bond length and bond angles, respectively.
A total of 109 ordered solvent molecules were
localized in the structure. This improved structure
at 2.3A°
led to an understanding of themechanism of
inhibition of the protein against a-chymotrypsin.An
analysis of this higher resolution structure also
helped us to predict the location of the second
reactive site of the protein, about which no previous
biochemical information was available. The inhibitor
structure is spherical and has twelve antiparallel
b-strands with connecting loops arranged
in a characteristic b-trefoil fold common to other
homologous serine protease inhibitors in the Kunitz
(STI) family as well as to some non homologous
functionally unrelated proteins. Awide variation in
the surface loop regions is seen in the latter ones.
Proteins 1999;35:321–331. r 1999Wiley-Liss, Inc