Selective interaction of megalin with postsynaptic density-95 (PSD-95)-like membrane-associated guanylate kinase (MAGUK) proteins

Mårten Larsson, Göran Hjälm, Amos M. Sakwe, Åke Engström, Anna Stina Höglund, Erik Larsson, Robert C. Robinson, Christian Sundberg, Lars Rask

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Megalin is an integral membrane receptor belonging to the low-density lipoprotein receptor family. In addition to its role as an endocytotic receptor, megalin has also been proposed to have signalling functions. Using interaction cloning in yeast, we identified the membrane-associated guanylate kinase family member postsynaptic density-95 (PSD-95) as an interaction partner for megalin. PSD-95 and a truncated version of megalin were co-immunoprecipitated from HEK-293 cell lysates overexpressing the two proteins, which confirmed the interaction. The two proteins were found to be co-localized in these cells by confocal microscopy. Immunocytochemical studies showed that cells in the parathyroid, proximal tubuli of the kidney and placenta express both megalin and PSD-95. We found that the interaction between the two proteins is mediated by the binding of the C-terminus of megalin, which has a type I PSD-95/Drosophila discs-large/zona occludens 1 (PDZ)-binding motif, to the PDZ2 domain of PSD-95. The PSD-95-like membrane-associated guanylate kinase ('MAGUK') family contains three additional members: PSD-93, synapse-associated protein 97 (SAP97) and SAP102. We detected these proteins, apart from SAP102, in parathyroid chief cells, a cell type having a marked expression of megalin. The PDZ2 domains of PSD-93 and SAP102 were also shown to interact with megalin, whereas no interaction was detected for SAP97. The SAP97 PDZ2 domain differed at four positions from the other members of the PSD-95 subfamily. One of these residues was Thr389, located in the αB-helix and part of the hydrophobic pocket of the PDZ2 domain. Surface plasmon resonance experiments revealed that mutation of SAP97 Thr389 to alanine, as with the other PSD-95-like membrane-associated guanylate kinases, induced binding to megalin.

Original languageEnglish
Pages (from-to)381-391
Number of pages11
JournalBiochemical Journal
Volume373
Issue number2
DOIs
Publication statusPublished - Jul 15 2003
Externally publishedYes

Fingerprint

Low Density Lipoprotein Receptor-Related Protein-2
Guanylate Kinases
Post-Synaptic Density
Proteins
Surface Plasmon Resonance
Confocal microscopy
LDL Receptors
HEK293 Cells
Cloning
Herpes Zoster
Surface plasmon resonance
Confocal Microscopy
Alanine
Yeast
Placenta
Drosophila
Organism Cloning
Yeasts

Keywords

  • Megalin
  • Membrane-associated guanylate kinase (MAGUK)
  • Postsynaptic density-95 (PSD-95)
  • PSD-95/Drosophila discs-large/zona occludens 1 (PDZ) domain
  • Synapse-associated protein 97 (SAP97)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Larsson, M., Hjälm, G., Sakwe, A. M., Engström, Å., Höglund, A. S., Larsson, E., ... Rask, L. (2003). Selective interaction of megalin with postsynaptic density-95 (PSD-95)-like membrane-associated guanylate kinase (MAGUK) proteins. Biochemical Journal, 373(2), 381-391. https://doi.org/10.1042/BJ20021958

Selective interaction of megalin with postsynaptic density-95 (PSD-95)-like membrane-associated guanylate kinase (MAGUK) proteins. / Larsson, Mårten; Hjälm, Göran; Sakwe, Amos M.; Engström, Åke; Höglund, Anna Stina; Larsson, Erik; Robinson, Robert C.; Sundberg, Christian; Rask, Lars.

In: Biochemical Journal, Vol. 373, No. 2, 15.07.2003, p. 381-391.

Research output: Contribution to journalArticle

Larsson, M, Hjälm, G, Sakwe, AM, Engström, Å, Höglund, AS, Larsson, E, Robinson, RC, Sundberg, C & Rask, L 2003, 'Selective interaction of megalin with postsynaptic density-95 (PSD-95)-like membrane-associated guanylate kinase (MAGUK) proteins', Biochemical Journal, vol. 373, no. 2, pp. 381-391. https://doi.org/10.1042/BJ20021958
Larsson, Mårten ; Hjälm, Göran ; Sakwe, Amos M. ; Engström, Åke ; Höglund, Anna Stina ; Larsson, Erik ; Robinson, Robert C. ; Sundberg, Christian ; Rask, Lars. / Selective interaction of megalin with postsynaptic density-95 (PSD-95)-like membrane-associated guanylate kinase (MAGUK) proteins. In: Biochemical Journal. 2003 ; Vol. 373, No. 2. pp. 381-391.
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