University of Kentucky

Poster Title

Enhanced Expression of Group V and X Secretory Phosphlipase A2: Two Enzymes Implicated in Cardiovascular Disease

Institution

University of Kentucky

Abstract

Cardiovascular disease is the leading cause of death in Kentucky. Two enzymes, Group V and Group X secretory phospolipase A2 (sPLA2), are thought to play a role in atherosclerosis, the most common cause of heart disease. However, investigation of their role in heart disease is difficult because these proteins are present in low amounts in tissues. The goal of this project is to develop methods to express and purify sufficient amounts of the enzymes for future studies. Many research groups have shown that a baculovirus expression system is often effective for producing mammalian proteins in insect cells. Baculovirus is an insect virus that naturally infects moths and butterflies. Recently, a novel system that produces higher baculovirus expression has been developed at the University of Kentucky. We determined whether this system could be used to express Group V and Group X sPLA2. We constructed two baculoviruses for each enzyme, one with and one without a fused protein sequence that was added to facilitate enzyme purification. Results will be presented to demonstrate the feasibility of our approach for generalizing pure, enzymatically active, Group V and Group X sPLA2.

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Enhanced Expression of Group V and X Secretory Phosphlipase A2: Two Enzymes Implicated in Cardiovascular Disease

Cardiovascular disease is the leading cause of death in Kentucky. Two enzymes, Group V and Group X secretory phospolipase A2 (sPLA2), are thought to play a role in atherosclerosis, the most common cause of heart disease. However, investigation of their role in heart disease is difficult because these proteins are present in low amounts in tissues. The goal of this project is to develop methods to express and purify sufficient amounts of the enzymes for future studies. Many research groups have shown that a baculovirus expression system is often effective for producing mammalian proteins in insect cells. Baculovirus is an insect virus that naturally infects moths and butterflies. Recently, a novel system that produces higher baculovirus expression has been developed at the University of Kentucky. We determined whether this system could be used to express Group V and Group X sPLA2. We constructed two baculoviruses for each enzyme, one with and one without a fused protein sequence that was added to facilitate enzyme purification. Results will be presented to demonstrate the feasibility of our approach for generalizing pure, enzymatically active, Group V and Group X sPLA2.