title: Monitoring collagen synthesis in fibroblasts using fluorescently labeled tRNA pairs.
abstract: There is a critical need for techniques that directly monitor protein synthesis within cells isolated from normal and diseased tissue. Fibrotic disease, for which there is no drug treatment, is characterized by the overexpression of collagens. Here, we use a bioinformatics approach to identify a pair of glycine and proline isoacceptor tRNAs as being specific for the decoding of collagen mRNAs, leading to development of a FRET-based approach, dicodon monitoring of protein synthesis (DiCoMPS), that directly monitors the synthesis of collagen. DiCoMPS aimed at detecting collagen synthesis will be helpful in identifying novel anti-fibrotic compounds in cells derived from patients with fibrosis of any etiology, and, suitably adapted, should be widely applicable in monitoring the synthesis of other proteins in cells.
authors.affiliation: Department of Physiology and Pharmacology, University of Western Ontario, London, Ontario, Canada; Department of Burns and Cutaneous Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, China.
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Liu
authors.forename: Macarena
authors.initials: M
authors.lastname: Pampillo
authors.forename: Fen
authors.initials: F
authors.lastname: Guo
authors.forename: Shangxi
authors.initials: S
authors.lastname: Liu
authors.forename: Barry S
authors.initials: BS
authors.lastname: Cooperman
authors.forename: Ian
authors.initials: I
authors.lastname: Farrell
authors.forename: Dvir
authors.initials: D
authors.lastname: Dahary
authors.forename: Bing S
authors.initials: BS
authors.lastname: Gan
authors.forename: David B
authors.initials: DB
authors.lastname: O'Gorman
authors.forename: Zeev
authors.initials: Z
authors.lastname: Smilansky
authors.forename: Andy V
authors.initials: AV
authors.lastname: Babwah
authors.forename: Andrew
authors.initials: A
authors.lastname: Leask
mesh_terms: D000818:Animals; D002232:Carbocyanines / Q000378:metabolism; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D005355:Fibrosis; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D051059:PTEN Phosphohydrolase / Q000172:deficiency / Q000235:genetics; D012353:RNA, Transfer, Gly / Q000235:genetics / Q000378:metabolism*; D012361:RNA, Transfer, Pro / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; D012353:RNA, Transfer, Gly; D012361:RNA, Transfer, Pro; C089187:cyanine dye 3; C085321:cyanine dye 5; D003094:Collagen; D051059:PTEN Phosphohydrolase; C487757:Pten protein, mouse