title: Fluorogenic label to quantify the cytosolic delivery of macromolecules.
abstract: The delivery of a macromolecule to the cytosol of human cells is assessed by using a pendant di-O-glycosylated derivative of fluorescein. Its fluorescence is unmasked by Escherichia coliβ-galactosidase installed in the cytosol. Background is diminished by using RNAi to suppress the expression of GLB1, which encodes a lysosomal β-galactosidase. This strategy was used to quantify the cytosolic entry of a highly cationic protein, ribonuclease A.
authors: Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706, USA.
authors: Tzu-Yuan
authors: TY
authors: Chao
authors: Ronald T
authors: RT
authors: Raines
mesh_terms: D029968:Escherichia coli Proteins / Q000096:biosynthesis; D005452:Fluoresceins / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D005697:Galactosides / Q000737:chemistry* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000235:genetics; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D029968:Escherichia coli Proteins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005697:Galactosides; D034741:RNA, Small Interfering; C038848:fluorescein-digalactoside; D012259:Ribonuclease, Pancreatic; D001616:beta-Galactosidase