 |
Shelley's Project Page - RISE Summer 2008 |
 |
Intern: Shelley Esakoff, Biochemistry, University of California, Santa Barbara
Mentor: Jen Getz
Faculty Supervisor: Patrick Daugherty
Department: Chemical Engineering |
ENGINEERING HIGH STABILITY AND HIGH AFFINITY THROMBIN-INHIBITING PEPTIDES
We are designing innovative high stability protein therapeutics to bind thrombin with high specificity and affinity
with the potential application of treating cardiovascular disease. Cardiovascular disease is the number one cause of
death in America with over 80 million fatalities a year. Many cardiovascular treatments target thrombin, a serine
protease that catalyzes the conversion of fibrinogen to fibrin, which is needed to form a stable blood clot. Current
treatments for cardiovascular disease, such as heparin, suffer from major adverse effects including excessive
bleeding and immunogenicity. Proteins are potentially effective pharmaceuticals due to their ability to achieve high
stability and high affinity. Craik et al. showed that the kalata B1 (KB1) scaffold found in the plant species
Oldenlandia affinis is robust in the presence of 8 M urea, temperatures up to 100°C and incubation with intestinal
proteases. Previously, peptides within the KB1 scaffold were isolated that bind thrombin using bacterial surface
display. A fluorescence resonance energy transfer (FRET) assay was then designed to determine the inhibitory effect
of the peptides on thrombin’s proteolytic activity. First the substrate’s affinity for thrombin needed to be determined
in order to characterize thrombin inhibition. Preliminary results have shown that the substrate’s affinity for thrombin
is approximately 50 µM.
Return to the RISE 2008 project list
