Abstract
Abstract: :
Purpose:Clone and sequence the phospholipases A2 (PLA2) expressed by human retinal pigmented epithelial (RPE) cells. Methods: Primers of different PLA2 were assayed by RT-PCR using RNA extracted from RPE prepared from human eye donors to specifically amplify the transcript of these proteins. The PCR products were then cloned and their identity was confirmed using NCBI data bank. Results:Human RPE cells express secreted PLA2s of group IIA and group V as well as the cytosolic PLA2 gamma which is a member of the group IV PLA2. Conclusion: RPE cells express three known phospholipases A2, i.e. two different secreted PLA2 (sPLA2) and one cytosolic PLA2 (cPLA2). Indeed, the sequence of both group V sPLA2 and cPLA2 gamma corresponds to the published sequences of these enzymes. However, given that we could not obtain the sequence of group IIA sPLA2 downstream of its catalytic site using different primers designed to amplified the full sequence of this enzyme, we conclude that human RPE cells express an isoform of group IIA sPLA2.
Keywords: 567 retinal pigment epithelium • 417 gene/expression • 308 age-related macular degeneration