title: PDZK1: I. a major scaffolder in brush borders of proximal tubular cells.
abstract: BACKGROUND
  In proximal tubular cells, PDZK1 (NaPi-Cap1) has been implicated in apical expression of the Na+-dependent phosphate cotransporter (NaPi-IIa) via interaction with its C-terminus. PDZK1 represents a multidomain protein consisting of four PDZ domains and thus is believed to have a broader specificity besides NaPi-IIa.
  
  
  METHODS
  We subjected single PDZ domains derived from PDZK1 either to yeast two-hybrid screens or yeast trap assays. Different pull-down assays and blot overlays were applied to corroborate the PDZK1-mediated interactions in vitro. Co-localization of interacting proteins with PDZK1 in proximal tubular cells was assessed by immunohistochemistry.
  
  
  RESULTS
  In the yeast screens, the most abundant candidate protein to interact with PDZK1 was the membrane-associated protein of 17 kD (MAP17). Besides MAP17, C-terminal parts of following transporters were also identified: NaPi-IIa, solute carrier SLC17A1 (NaPi-I), Na+/H+ exchanger (NHE-3), organic cation transporter (OCTN1), chloride-formate exchanger (CFEX), and urate-anion exchanger (URAT1). In addition, other regulatory factors were found among the clones, such as a protein kinase A (PKA)-anchoring protein (D-AKAP2) and N+/H+ exchanger regulator factor (NHERF-1). All interactions of itemized proteins with PDZK1 were affirmed by in vitro techniques. Apart from PDZK1, strong in vitro interactions of NHERF-1 were also observed with the solute transporters (excluding MAP17) and D-AKAP2. All identified proteins were immunolocalized in proximal tubular cells, wherein all membrane proteins co-localized with PDZK1 in brush borders.
  
  
  CONCLUSIONS
  We hypothesize that PDZK1 and NHERF-1 establish an extended network beneath the apical membrane to which membrane proteins and regulatory components are anchored.
authors.affiliation: Department of Physiology and Biochemistry, University of Zürich, Zürich, Switzerland.
authors.forename: Serge M
authors.initials: SM
authors.lastname: Gisler
authors.forename: Sandra
authors.initials: S
authors.lastname: Pribanic
authors.forename: Desa
authors.initials: D
authors.lastname: Bacic
authors.forename: Patrik
authors.initials: P
authors.lastname: Forrer
authors.forename: Andrea
authors.initials: A
authors.lastname: Gantenbein
authors.forename: Luc A
authors.initials: LA
authors.lastname: Sabourin
authors.forename: Akira
authors.initials: A
authors.lastname: Tsuji
authors.forename: Zhuo-Shen
authors.initials: ZS
authors.lastname: Zhao
authors.forename: Edward
authors.initials: E
authors.lastname: Manser
authors.forename: Jürg
authors.initials: J
authors.lastname: Biber
authors.forename: Heini
authors.initials: H
authors.lastname: Murer
mesh_terms: D000818:Animals; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D007150:Immunohistochemistry; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008871:Microvilli / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D017923:Sodium-Hydrogen Exchangers; D020798:Two-Hybrid System Techniques; D015003:Yeasts
chemical_list: D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D010750:Phosphoproteins; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor