Presentation
Effect of cyclooxygenase-2 glycosylation on downstream prostanoid synthesis pathways
National Conference on Undergraduate Research, Weber State University
(2012)
Abstract
Cyclooxygenase-2 (COX-2) is an enzyme that catalyzes the rate-limiting step in the prostanoid synthesis pathway, which plays an important role in a variety of physiological and pathophysiological processes, including inflammation and cancer. COX-2 exists as two major glycoforms of 72 and 74 kDa, the latter resulting from an additional oligosaccharide chain at amino acid residue Asn580. The purpose of this study is to determine if this additional glycosylation site affects the downstream production of prostaglandins. COS-1 cells were transiently transfected with either the wild- type or Asn580-mutant COX-2 gene. After the addition of the COX-2 substrate arachidonic acid to the cells, media was collected and acidified to pH 3.5 with 0.5 M citric acid. Prostaglandins were then extracted with ethyl acetate and dried under N2. After resuspension in ethyl alcohol, the prostaglandin samples were spotted onto silica- coated thin- layer chromatography plates, which were resolved with chloroform: methanol: acetic acid: water (90: 8: 1: 1) and visualized with 3% cuprous acetate and 15% phosphoric acid. We predict that we will find differences in prostaglandin profiles between the cells expressing both the 72 and 74 kDa glycoforms and the cells unable to express the 74 kDa glycoform. If our prediction is correct, this would suggest that the additional glycosylation of COX-2 at Asn580 affects its interactions with downstream prostanoid synthases.
Disciplines
Publication Date
April, 2012
Location
Ogden, UT
Citation Information
Josephine Stipe, Mary B. Sevigny and Maria G Carranza. "Effect of cyclooxygenase-2 glycosylation on downstream prostanoid synthesis pathways" National Conference on Undergraduate Research, Weber State University (2012) Available at: http://works.bepress.com/mary-sevigny/17/