Visual cycle retinoid processing proteins are present in HEK293S cells.
In HEK293S cells expressing opsin, rhodopsin regenerates on addition of all-trans retinol. This study was to determine if key proteins in the retinal pigment epithelium (RPE) are present in these cells. Cellular retinoid binding protein, cellular retinoic-acid binding protein, RPE65, caveolin-1-alpha- and -beta-isoforms, interphotoreceptor retinoid binding protein, and 11-cis retinol dehydrogenase, but not lecithin:retinol acyltransferase (LRAT), were identified by Western blot analysis. LRAT transcripts were found by RT-PCR and Southern blot analysis. Small interference RNA specific to LRAT reduced ester formation, confirming that the enzyme is present. Therefore, HEK293S cells contain the functional components of the retinoid cycle found in the RPE.
Chen
Yumei
Y
Department of Ophthalmology, Medical University of South Carolina, 167 Ashley Avenue, 29425, Charleston, SC, USA
Moiseyev
Gennadiy
G
Wu
Bill X
BX
Ma
Jian-xing
JX
Crouch
Rosalie K
RK
D000217:Acyltransferases / Q000032:analysis; D015139:Blotting, Southern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D002352:Carrier Proteins; D051242:Caveolin 1; D022461:Caveolins / Q000032:analysis; D002478:Cells, Cultured / Q000737:chemistry*; D005136:Eye Proteins; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000166:cytology*; D010857:Pigment Epithelium of Eye / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods; D011506:Proteins / Q000032:analysis; D012313:RNA / Q000032:analysis; D018168:Receptors, Retinoic Acid / Q000032:analysis; D012176:Retinoids / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D019745:cis-trans-Isomerases; D000099214:Retinol-Binding Proteins, Interstitial; D000099178:Retinoid Isomerohydrolase
D000217:Acyltransferases; D002352:Carrier Proteins; D051242:Caveolin 1; D022461:Caveolins; D005136:Eye Proteins; D011506:Proteins; D012313:RNA; D018168:Receptors, Retinoic Acid; D012176:Retinoids; D012177:Retinol-Binding Proteins; D019745:cis-trans-Isomerases; D000099214:Retinol-Binding Proteins, Interstitial; D000099178:Retinoid Isomerohydrolase; C495540:CAV1 protein, human; C056942:lecithin-retinol acyltransferase