Scorpion venom may help treat pancreatitis

Researchers at North Carolina State University and East Carolina University have
gained insight into scorpion venom’s effects on the ability of certain cells to release
critical components - a finding that may prove useful in understanding diseases like
pancreatitis or in targeted drug delivery.

A common result of scorpion stings, pancreatitis is an inflammation of the pancreas.

ECU microbiologist Dr. Paul Fletcher believed that scorpion venom might be used as a
way to discover how pancreatitis occurs - to see which cellular processes are affected at
the onset of the disease.

Fletcher pinpointed a protein production system found in the pancreas that seemed to be
targeted by the venom of the Brazilian scorpion Tityus serrulatus and then contacted NC
State physicist Dr. Keith Weninger, who had studied that particular protein system.

"This particular protein system has special emphasis at two places in the body - the
pancreas and the nervous system," Weninger says. "In the pancreas, it is involved in the
release of proteins through the membrane of a cell."

The pancreas specializes in releasing two kinds of proteins using separate cells:
digestive enzymes that go into the small intestine and insulin and its relatives that go into
the bloodstream, yet this same release mechanism is important in all of our cells for
many processes.

Cells move components in and out through a process called vesicle fusion. The vesicle
is a tiny, bubble-like chamber inside the cell that contains the substance to be moved,
stored and released - in this case, proteins like enzymes or hormones. The vesicle is
moved through the cell and attaches to the exterior membrane, where the vesicle acts
like an airlock in a spaceship, allowing the cell membrane to open and release the
proteins without disturbing the rest of the cell’s contents. The proteins that aid in this
process are known as Vesicle Associated Membrane Proteins, or VAMPs.

Weninger provided Fletcher with two different VAMP proteins found in the pancreas,
VAMP2 and VAMP8. They were engineered to remove the membrane attachments so
they could be more easily used for experiments outside cells and tissues. Fletcher’s
team demonstrated that the scorpion venom attacked the VAMP proteins, cutting them in
one place and eliminating the vesicle’s ability to transport its protein cargo out of the cell.i
Scorpion Venom Provides Clues to Cause, Treatment of
Pancreatitis

March 29, 2010 Tracey Peake

A Brazilian scorpion has provided researchers at North Carolina State University and
East Carolina University insight into venom’s effects on the ability of certain cells to
release critical components. The findings may prove useful in understanding diseases
like pancreatitis or in targeted drug delivery.

A common result of scorpion stings, pancreatitis is an inflammation of the pancreas.

ECU microbiologist Dr. Paul Fletcher believed that scorpion venom might be used as a
way to discover how pancreatitis occurs – to see which cellular processes are affected at
the onset of the disease. Fletcher pinpointed a protein production system found in the
pancreas that seemed to be targeted by the venom of the Brazilian scorpion Tityus
serrulatus and then contacted NC State physicist Dr. Keith Weninger, who had studied
that particular protein system.

“This particular protein system has special emphasis at two places in the body – the
pancreas and the nervous system,” Weninger says. “In the pancreas, it is involved in the
release of proteins through the membrane of a cell.” The pancreas specializes in
releasing two kinds of proteins using separate cells: digestive enzymes that go into the
small intestine and insulin and its relatives that go into the bloodstream, yet this same
release mechanism is important in all of our cells for many processes.

Cells move components in and out through a process called vesicle fusion. The vesicle
is a tiny, bubble-like chamber inside the cell that contains the substance to be moved,
stored and released – in this case, proteins like enzymes or hormones. The vesicle is
moved through the cell and attaches to the exterior membrane, where the vesicle acts
like an airlock in a spaceship, allowing the cell membrane to open and release the
proteins without disturbing the rest of the cell’s contents. The proteins that aid in this
process are known as Vesicle Associated Membrane Proteins, or VAMPs.

Weninger provided Fletcher with two different VAMP proteins found in the pancreas,
VAMP2 and VAMP8. They were engineered to remove the membrane attachments so
they could be more easily used for experiments outside cells and tissues. Fletcher’s
team demonstrated that the scorpion venom attacked the VAMP proteins, cutting them in
one place and eliminating the vesicle’s ability to transport its protein cargo out of the cell.

The results were published in the March 5 issue of the Journal of Biological Chemistry.
“We found that a particular enzyme in the scorpion’s venom removes a peptide, or small
protein, that allows the vesicle to fuse with the cell membrane,” Fletcher says. “If you
remove a pancreatic cell’s ability to absorb or release components, you end up with
pancreatitis.”

“Viruses often exploit the same mechanism of vesicle fusion, but in reverse, in order to
invade cells and replicate,” Weninger adds. “This work furthers our understanding of a
basic cellular process and may lead to treatments for viruses and advances in
treatments like chemotherapy, by allowing targeted drug delivery only to cancer cells.”

The Department of Physics is part of NC State University’s College of Physical and

Mathematical Sciences.

-peake-

Note to editors: An abstract of the paper follows.

“Vesicle-associated Membrane Protein (VAMP) Cleavage by a New Metalloprotease

from the Brazilian Scorpion Tityus serrulatus*”
Authors: Paul L. Fletcher, Jr., Maryann D. Fletcher, East Carolina University; Keith
Weninger, Trevor E. Anderson, North Carolina State University; and Brian M. Martin,
National Institutes of Health
Published: March 5, 2010, in Journal of Biological Chemistry; VOL. 285, NO. 10, pp.
7405–7416

Abstract: We present evidence that venom from the Brazilian scorpion Tityus
serrulatus and a purified fraction selectively cleave essential SNARE proteins within
exocrine pancreatic tissue. Western blotting for vesicle-associated membrane protein
type v-SNARE proteins (or synaptobrevins) reveals characteristic alterations to
venom-treated excised pancreatic lobules in vitro. Immunocytochemistry by electron
microscopy confirms both the SNARE identity as VAMP2 and the proteolysis of VAMP2
as a marked decrease in secondary antibody-conjugated colloidal gold particles that are
predominantly associated with mature zymogen granules. Studies with recombinant
SNARE proteins were used to determine the specific cleavage site in VAMP2 and the
susceptibility of VAMP8 (endobrevin). The VAMP2 cleavage site is between the
transmembrane anchor and the SNARE motif that assembles into the ternary SNARE
complex. Inclusion of divalent chelating agents (EDTA) with fraction v, an otherwise
active purified component from venom, eliminates SNARE proteolysis, suggesting the
active protein is a metalloprotease. The unique cleavages of VAMP2 and VAMP8 may
be linked to pancreatitis that develops following scorpion envenomation as both of these
v-SNARE proteins are associated with zymogen granule membranes in pancreatic
acinar cells. We have isolated antarease, a metalloprotease from fraction v that cleaves
VAMP2, and report its amino acid sequence.
i

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