  A novel uracil-DNA degrading protein factor (termed UDE) was identified in
Drosophila melanogaster with no significant structural and functional homology
to other uracil-DNA binding or processing factors. Determination of the 3D
structure of UDE will be a true breakthrough in description of the molecular
mechanism of action of UDE catalysis, as well as in general uracil-recognition
and nuclease action. The revolutionary ESPRIT technology was applied to the
novel protein UDE to overcome problems in identifying soluble expressing
constructs given the absence of precise information on domain content and
arrangement. Nine specimen from the created numerous truncated constructs of
UDE were choosen to dechiper structural and functional relationships. VUVCD
with neural network was performed to define the secondary structure content and
location of UDE and its truncated variants. The quantitative analysis
demonstrated exclusive {\alpha}-helical content for the full-length protein,
which is preserved in the truncated constructs. Partition of {\alpha}-helical
boundles comparison with the truncated protein segments denoted new domain
boundaries which differ from the conserved motifs determined by sequence-based
alignment of UDE homologues in pupating insects. Here we demonstrate that
combination of ESPRIT and VUVCD with NN provided significant structural
description of UDE and resulted useful truncated constructs for further
detailed functional studies.
