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R28
LABORATORY OF BARBARA FURIE AND BRUCE FURIE
|Pub/ications
Brown, M., B. Hambe, B.
Furie, B. C. Furie, J.
Stenflo, and L M
Stenberg. 2002. Detection
of vitamin K-dependent
in proteins venoms with a
monoclonal antibody
specific for p-
carboxyglutamic acid,
lexicon 40: 447-453,
Czerwiec, E., G. S. Begley,
J. Stenflo, K Taylor, B. C.
Furie, and B Furie. 2002.
Structural similarity and
functional differences
between invertebrate and
vertebrate carboxylases:
expression and characterization
of recombinant
vitamin K-dependent (Jglutamyl
carboxylase from
Conus textile. ur. J.
Biochem. 269:6162-6172.
P-Carboxyglutamic acid is a calcium-binding amino acid that is found
in the conopeptides of the predatory marine cone snail, Conus.
This
laboratory has been investigating the biosynthesis of this amino acid in
Conus and the structural role of p-carboxyglutamic acid in the
conopeptides.
This satellite laboratory relates closely to the main
laboratory, the Center for Hemostasis and Thrombosis Research, on the
Harvard Medical School campus in Boston, whose main focus is the
[Staff
synthesis and function of p-carboxyglutamic acid in blood clotting
proteins and the role of vitamin K.
Until recently, the marine cone snail had been the sole invertebrate
known to synthesize the vitamin K-dependent amino acid, p--
carboxyglutamic acid (Gla), but the work of this laboratory and others
has shown that this synthetic pathway has been preserved in most animal phyla.
ADJUNCT SCIENTISTS
Barbara C. Furie. Harvard
Medical School
Bruce Furie, Harvard
Medical School
Alan Rigby, Harvard
Medical School
VISITING SCIENTIST
Johan Stenflo, University of Lund
STAFF SCIENTIST II
Eva Czerwiec
The cone snail
produces neurotoxic conopeptides, some rich in Gla, which it
injects into its prey to immobilize
it. To examine the biosynthetic pathway for Gla, we have studied the Conus carboxylase which
converts glutamic acid to p-carboxyglutamic acid in the presence of vitamin K. We examined
the diversity of animal species that maintain vitamin K-dependent carboxylation to generate
Gla. We have cloned full length carboxylase from the beluga whale (De/ph/napterus /eucas), the
toadfish (Opsanus tau), and the cone snail (Conus text//e) to compare these structures to the
known bovine, human, rat, and mouse cDNA sequences.
In addition, we have partially cloned
the carboxylase gene from chicken (Ga//us gallus), hagfish (Myxine g/ut/nosa), and horseshoe
crab (L/mu/us po/yphemus).
In addition, the Drosophi/a genome contains the p-carboxylase
gene. The predicted amino acid sequence of the carboxylase cDNA from Conus textile shows
most regions are nearly identical to the mammalian sequence, and that there is about 40%
sequence similarity.
This protein has been expressed, and the recombinant enzyme identified
as a carboxylase and epoxidase. These results demonstrate the broad distribution of the vitamin
Cone snails, Volker Steger
-dependent carboxylase gene, including a highly conserved motif that is
likely critical for
enzyme function. The vitamin K-dependent biosynthesis of Gla is a highly conserved function in
the animal kingdom and we are now searching for a novel Gla containing protein that is critical
for survival of animal species.