title: The C-terminal domain controls the mobility of Crumbs 3 isoforms.
abstract: The physiological function of epithelia depends on an asymmetric distribution of their membrane domains. Polarity proteins play a crucial role for distribution processes, however, little is known about their mobility in epithelial cells. In this study, we analyzed the intracellular and plasma-membrane-associated mobility of fluorescence-labeled Crb3A and Crb3B. Both variants belong to the Crumbs protein family, which control size and identity of apical membranes in epithelial cells. Fluorescence recovery after photo-bleaching measurements revealed different mobilities for the two Crb3 variants. They also differentially affected mobility and localization of the Pals1/Mpp5 protein, which binds to Crb3A but not to Crb3B. In addition, tracking of intracellular vesicles indicated that Crb3A containing vesicles are slightly more immobile than Crb3B ones. Taken together, our data revealed different intracellular mobility patterns for Crb3A and Crb3B.
authors.affiliation: Internal Medicine D, Molecular Nephrology, University Hospital of Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: Ivona
authors.initials: I
authors.lastname: Djuric
authors.affiliation: Institute of Physical and Theoretical Chemistry, Rheinische Friedrich-Wilhelms University Bonn, Wegelerstraße 12, 53115 Bonn, Germany.
authors.forename: Jan Peter
authors.initials: JP
authors.lastname: Siebrasse
authors.affiliation: Internal Medicine D, Molecular Nephrology, University Hospital of Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: Ulf
authors.initials: U
authors.lastname: Schulze
authors.affiliation: Internal Medicine D, Molecular Nephrology, University Hospital of Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Granado
authors.affiliation: Internal Medicine D, Molecular Nephrology, University Hospital of Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: Marc A
authors.initials: MA
authors.lastname: Schlüter
authors.affiliation: Institute of Physical and Theoretical Chemistry, Rheinische Friedrich-Wilhelms University Bonn, Wegelerstraße 12, 53115 Bonn, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Kubitscheck
authors.affiliation: Internal Medicine D, Molecular Nephrology, University Hospital of Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany. Electronic address: hermann.pavenstaedt@ukmuenster.de.
authors.forename: Hermann
authors.initials: H
authors.lastname: Pavenstädt
authors.affiliation: Internal Medicine D, Molecular Nephrology, University Hospital of Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany. Electronic address: weidet@uni-muenster.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Weide
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D002461:Cell Line, Transformed; D036681:Fluorescence Recovery After Photobleaching; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009703:Nucleoside-Phosphate Kinase / Q000235:genetics / Q000378:metabolism; D050199:Podocytes / Q000166:cytology / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques
chemical_list: C472002:CRB3 protein, human; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D009703:Nucleoside-Phosphate Kinase; C501307:MPP5 protein, human
keywords: Cell polarity; Crb; Crb3; Crumbs; FRAP; Mobility; Pals1; Single particle tracking