Prostaglandin receptor EP1-mediated differential degradation of cyclooxygenases involves a specific lysine residue

•COX-1 and COX-2 differ in their sensitivity to EP1 receptor-aided degradation.•Mutation of a putative target of ubiquitination on COX-2 (K432R) decreases its sensitivity to EP1.•Insertion of a parallel ubiquitination site (H446K′) into COX-1 increases its sensitivity to EP1.•Distinctive ubiquitination may regulate the stability of COX isoforms. The cyclooxygenase (COX) enzyme isoforms COX-1 and COX-2 catalyze the main step in the generation of prostanoids that mediate major physiological functions. Whereas COX-1 is a ubiquitously expressed stable protein, COX-2 is transiently upregulated in many pathologies and is often associated with a poor prognostic outcome. We have recently shown that an interaction of COX-2 with the prostaglandin EP1 receptor accelerates its degradation via a mechanism that augments its level of ubiquitination. Here we show that the sensitivity of both COX-1 and COX-2 to EP1 is altered upon modification of one lysine residue. A point mutation of lysine to-arginine in position 432 of COX-2 (K432R) yields an enzyme with decreased sensitivity to EP1-mediated degradation. In contrast, insertion of a putative ubiquitination site into the corresponding position of COX-1 (H446K′) yields an enzyme with higher levels of ubiquitination and reduced expression. Furthermore, compared to wild type COX-1, H446K′ is significantly more sensitive to downregulation by EP1. Together these data suggest that distinctive ubiquitination of COX-1 and COX-2 may be responsible for their different sensitivity to EP1-mediated degradation.