Role of the N-terminal transmembrane region of the multidrug resistance protein MRP2 in routing to the apical membrane in MDCKII cells

Sara B. Mateus Fernández, Zsolt Holló, Andras Kern, E. Bakos, Paul A. Fischer, Piet Borst, Raymond Evers

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Abstract

In polarized cells, the multidrug resistance protein MRP2 is localized in the apical plasma membrane, whereas MRP1, another multidrug resistance protein (MRP) family member, is localized in the basolateral membrane. MRP1 and MRP2 are thought to contain an N-terminal region of five transmembrane segments (TMD0) coupled to 2 times six transmembrane segments via an intracellular loop (L0). We previously demonstrated for MRP1 that a mutant lacking TMD0 but still containing L0, called L0ΔMRP1, was functional and routed to the lateral plasma membrane. To investigate the role of the TMD0L0 region of MRP2 in routing to the apical membrane, we generated mutants similar to those made for MRP1. In contrast to L0ΔMRP1, L0ΔMRP2 was associated with an intracellular compartment, most likely endosomes. Co-expression with TMD0, however, resulted in apical localization of L0ΔMRP2 and transport activity. Uptake experiments with vesicles containing L0ΔMRP2 demonstrated that the molecule is able to transport LTC4. An MRP2 mutant without TMD0L0, ΔMRP2, was only core-glycosylated and localized intracellularly. Co-expression of ΔMRP2 with TMD0L0 resulted in an increased protein level of ΔMRP2, full glycosylation of the protein, routing to the apical membrane, and transport activity. Our results suggest that the TMD0 region is required for routing to or stable association with the apical membrane.

Original languageEnglish
Pages (from-to)31048-31055
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number34
DOIs
Publication statusPublished - Aug 23 2002

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Membranes
Cell membranes
Cell Membrane
P-Glycoproteins
Glycosylation
Leukotriene C4
Endosomes
Proteins
Association reactions
Molecules
multidrug resistance-associated protein 2
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Role of the N-terminal transmembrane region of the multidrug resistance protein MRP2 in routing to the apical membrane in MDCKII cells. / Mateus Fernández, Sara B.; Holló, Zsolt; Kern, Andras; Bakos, E.; Fischer, Paul A.; Borst, Piet; Evers, Raymond.

In: Journal of Biological Chemistry, Vol. 277, No. 34, 23.08.2002, p. 31048-31055.

Research output: Contribution to journalArticle

Mateus Fernández, Sara B. ; Holló, Zsolt ; Kern, Andras ; Bakos, E. ; Fischer, Paul A. ; Borst, Piet ; Evers, Raymond. / Role of the N-terminal transmembrane region of the multidrug resistance protein MRP2 in routing to the apical membrane in MDCKII cells. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 34. pp. 31048-31055.
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abstract = "In polarized cells, the multidrug resistance protein MRP2 is localized in the apical plasma membrane, whereas MRP1, another multidrug resistance protein (MRP) family member, is localized in the basolateral membrane. MRP1 and MRP2 are thought to contain an N-terminal region of five transmembrane segments (TMD0) coupled to 2 times six transmembrane segments via an intracellular loop (L0). We previously demonstrated for MRP1 that a mutant lacking TMD0 but still containing L0, called L0ΔMRP1, was functional and routed to the lateral plasma membrane. To investigate the role of the TMD0L0 region of MRP2 in routing to the apical membrane, we generated mutants similar to those made for MRP1. In contrast to L0ΔMRP1, L0ΔMRP2 was associated with an intracellular compartment, most likely endosomes. Co-expression with TMD0, however, resulted in apical localization of L0ΔMRP2 and transport activity. Uptake experiments with vesicles containing L0ΔMRP2 demonstrated that the molecule is able to transport LTC4. An MRP2 mutant without TMD0L0, ΔMRP2, was only core-glycosylated and localized intracellularly. Co-expression of ΔMRP2 with TMD0L0 resulted in an increased protein level of ΔMRP2, full glycosylation of the protein, routing to the apical membrane, and transport activity. Our results suggest that the TMD0 region is required for routing to or stable association with the apical membrane.",
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