Receptor-associated protein is important for normal processing of megalin in kidney proximal tubules

Henrik Birn; Henrik Vorum; Pierre J. Verroust; Søren K. Moestrup; Erik Ilsø Christensen

doi:10.1681/ASN.V112191

Receptor-associated protein is important for normal processing of megalin in kidney proximal tubules

Henrik Birn^*
, Henrik Vorum
, Pierre J. Verroust
, Søren K. Moestrup
, Erik Ilsø Christensen

^*Kontaktforfatter

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

91 Citationer (Scopus)

Abstract

The receptor-associated protein (RAP) has been identified as a chaperone regulating the expression and processing of the LDL receptor-related protein. RAP also binds to the related 600-kD multiligand endocytic receptor megalin expressed in many absorptive epithelia including renal proximal tubule. The present study examines the effect of RAP gene disruption on megalin expression and subcellular distribution in the proximal tubule as well as the effect on tubular protein reabsorption. It is shown that RAP is important for the normal expression and function of megalin. Megalin expression was reduced to approximately 23% estimated by immunoblotting and supported by immunocytochemistry and by the amount of megalin recovered by RAP affinity chromatography. Light-and electron microscope immunocytochemistry as well as analyses on separated membrane fractions showed significant changes in the subcellular distribution of megalin. A significant reduction in the normal brush border labelling was observed in association with increased labeling of rough endoplasmic reticulum and the smooth paramembranous endoplasmic reticulum along the basolateral membranes. RAP deficiency was associated with changes in urinary protein composition, enabling the identification of α- amylase as a new ligand for megalin. In addition, an increased excretion of vitamin D-binding protein, a recently identified ligand to megalin, was observed Supporting changes in tubular protein reabsorption. The present data show that RAP is of crucial importance for normal processing and function of megalin, suggesting a chaperone-like function of this protein in the kidney proximal tubule.

Originalsprog	Engelsk
Tidsskrift	Journal of the American Society of Nephrology
Vol/bind	11
Udgave nummer	2
Sider (fra-til)	191-202
Antal sider	12
ISSN	1046-6673
DOI	https://doi.org/10.1681/ASN.V112191
Status	Udgivet - feb. 2000
Udgivet eksternt	Ja

Bibliografisk note

Funding Information:
This work was supported by the Danish Medical Research Council, the University of Aarhus, the NOVO-Nordisk Foundation, the Leo Nielsen Foundation, the Ruth König-Petersen Foundation, Fonden til Lægevidenskabens Fremme, the Beckett Foundation, and the Bi-omembrane Research Center, University of Aarhus. The authors thank Drs. Jan Stagstad, Poul-Henning Jensen, and Anders Nykjær (Institute of Medical Biochemistry, University of Aarhus, Denmark) for the kind gift of the RAP knockout mice, the anti-RAP antibody, and the anti-mannose-6-phosphate receptor antibody, respectively. We also thank Dr. Søren Nielsen (Department of Cell Biology, University of Aarhus, Denmark) and Dr. Mark Knepper (Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, MD) for the donation of the anti-AQP-1 antibody. The skillful technical assistance of Mette Vistisen, Ann Vad Steffensen, Hanne Sidelmann, Inger Kristoffersen, and Albert Meier is greatly appreciated.

Funding Information:
This work was supported by the Danish Medical Research Council, the University of Aarhus, the NOVO-Nordisk Foundation, the Leo Nielsen Foundation, the Ruth K?nig-Petersen Foundation, Fonden til L?gevidenskabens Fremme, the Beckett Foundation, and the Biomembrane Research Center, University of Aarhus. The authors thank Drs. Jan Stagstad, Poul-Henning Jensen, and Anders Nykj?r (Institute of Medical Biochemistry, University of Aarhus, Denmark) for the kind gift of the RAP knockout mice, the anti-RAP antibody, and the anti-mannose-6-phosphate receptor antibody, respectively. We also thank Dr. S?ren Nielsen (Department of Cell Biology, University of Aarhus, Denmark) and Dr. Mark Knepper (Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, MD) for the donation of the anti-AQP-1 antibody. The skillful technical assistance of Mette Vistisen, Ann Vad Steffensen, Hanne Sidelmann, Inger Kristoffersen, and Albert Meier is greatly appreciated.

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10.1681/ASN.V112191

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@article{b97e66698a604535815702f12e8579ce,

title = "Receptor-associated protein is important for normal processing of megalin in kidney proximal tubules",

abstract = "The receptor-associated protein (RAP) has been identified as a chaperone regulating the expression and processing of the LDL receptor-related protein. RAP also binds to the related 600-kD multiligand endocytic receptor megalin expressed in many absorptive epithelia including renal proximal tubule. The present study examines the effect of RAP gene disruption on megalin expression and subcellular distribution in the proximal tubule as well as the effect on tubular protein reabsorption. It is shown that RAP is important for the normal expression and function of megalin. Megalin expression was reduced to approximately 23\% estimated by immunoblotting and supported by immunocytochemistry and by the amount of megalin recovered by RAP affinity chromatography. Light-and electron microscope immunocytochemistry as well as analyses on separated membrane fractions showed significant changes in the subcellular distribution of megalin. A significant reduction in the normal brush border labelling was observed in association with increased labeling of rough endoplasmic reticulum and the smooth paramembranous endoplasmic reticulum along the basolateral membranes. RAP deficiency was associated with changes in urinary protein composition, enabling the identification of α- amylase as a new ligand for megalin. In addition, an increased excretion of vitamin D-binding protein, a recently identified ligand to megalin, was observed Supporting changes in tubular protein reabsorption. The present data show that RAP is of crucial importance for normal processing and function of megalin, suggesting a chaperone-like function of this protein in the kidney proximal tubule.",

author = "Henrik Birn and Henrik Vorum and Verroust, \{Pierre J.\} and Moestrup, \{S{\o}ren K.\} and Christensen, \{Erik Ils{\o}\}",

note = "Funding Information: This work was supported by the Danish Medical Research Council, the University of Aarhus, the NOVO-Nordisk Foundation, the Leo Nielsen Foundation, the Ruth K{\"o}nig-Petersen Foundation, Fonden til L{\ae}gevidenskabens Fremme, the Beckett Foundation, and the Bi-omembrane Research Center, University of Aarhus. The authors thank Drs. Jan Stagstad, Poul-Henning Jensen, and Anders Nykj{\ae}r (Institute of Medical Biochemistry, University of Aarhus, Denmark) for the kind gift of the RAP knockout mice, the anti-RAP antibody, and the anti-mannose-6-phosphate receptor antibody, respectively. We also thank Dr. S{\o}ren Nielsen (Department of Cell Biology, University of Aarhus, Denmark) and Dr. Mark Knepper (Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, MD) for the donation of the anti-AQP-1 antibody. The skillful technical assistance of Mette Vistisen, Ann Vad Steffensen, Hanne Sidelmann, Inger Kristoffersen, and Albert Meier is greatly appreciated. Funding Information: This work was supported by the Danish Medical Research Council, the University of Aarhus, the NOVO-Nordisk Foundation, the Leo Nielsen Foundation, the Ruth K?nig-Petersen Foundation, Fonden til L?gevidenskabens Fremme, the Beckett Foundation, and the Biomembrane Research Center, University of Aarhus. The authors thank Drs. Jan Stagstad, Poul-Henning Jensen, and Anders Nykj?r (Institute of Medical Biochemistry, University of Aarhus, Denmark) for the kind gift of the RAP knockout mice, the anti-RAP antibody, and the anti-mannose-6-phosphate receptor antibody, respectively. We also thank Dr. S?ren Nielsen (Department of Cell Biology, University of Aarhus, Denmark) and Dr. Mark Knepper (Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, MD) for the donation of the anti-AQP-1 antibody. The skillful technical assistance of Mette Vistisen, Ann Vad Steffensen, Hanne Sidelmann, Inger Kristoffersen, and Albert Meier is greatly appreciated.",

year = "2000",

month = feb,

doi = "10.1681/ASN.V112191",

language = "English",

volume = "11",

pages = "191--202",

journal = "Journal of the American Society of Nephrology",

issn = "1046-6673",

publisher = "Wolters Kluwer",

number = "2",

}

TY - JOUR

T1 - Receptor-associated protein is important for normal processing of megalin in kidney proximal tubules

AU - Birn, Henrik

AU - Vorum, Henrik

AU - Verroust, Pierre J.

AU - Moestrup, Søren K.

AU - Christensen, Erik Ilsø

N1 - Funding Information: This work was supported by the Danish Medical Research Council, the University of Aarhus, the NOVO-Nordisk Foundation, the Leo Nielsen Foundation, the Ruth König-Petersen Foundation, Fonden til Lægevidenskabens Fremme, the Beckett Foundation, and the Bi-omembrane Research Center, University of Aarhus. The authors thank Drs. Jan Stagstad, Poul-Henning Jensen, and Anders Nykjær (Institute of Medical Biochemistry, University of Aarhus, Denmark) for the kind gift of the RAP knockout mice, the anti-RAP antibody, and the anti-mannose-6-phosphate receptor antibody, respectively. We also thank Dr. Søren Nielsen (Department of Cell Biology, University of Aarhus, Denmark) and Dr. Mark Knepper (Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, MD) for the donation of the anti-AQP-1 antibody. The skillful technical assistance of Mette Vistisen, Ann Vad Steffensen, Hanne Sidelmann, Inger Kristoffersen, and Albert Meier is greatly appreciated. Funding Information: This work was supported by the Danish Medical Research Council, the University of Aarhus, the NOVO-Nordisk Foundation, the Leo Nielsen Foundation, the Ruth K?nig-Petersen Foundation, Fonden til L?gevidenskabens Fremme, the Beckett Foundation, and the Biomembrane Research Center, University of Aarhus. The authors thank Drs. Jan Stagstad, Poul-Henning Jensen, and Anders Nykj?r (Institute of Medical Biochemistry, University of Aarhus, Denmark) for the kind gift of the RAP knockout mice, the anti-RAP antibody, and the anti-mannose-6-phosphate receptor antibody, respectively. We also thank Dr. S?ren Nielsen (Department of Cell Biology, University of Aarhus, Denmark) and Dr. Mark Knepper (Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, MD) for the donation of the anti-AQP-1 antibody. The skillful technical assistance of Mette Vistisen, Ann Vad Steffensen, Hanne Sidelmann, Inger Kristoffersen, and Albert Meier is greatly appreciated.

PY - 2000/2

Y1 - 2000/2

N2 - The receptor-associated protein (RAP) has been identified as a chaperone regulating the expression and processing of the LDL receptor-related protein. RAP also binds to the related 600-kD multiligand endocytic receptor megalin expressed in many absorptive epithelia including renal proximal tubule. The present study examines the effect of RAP gene disruption on megalin expression and subcellular distribution in the proximal tubule as well as the effect on tubular protein reabsorption. It is shown that RAP is important for the normal expression and function of megalin. Megalin expression was reduced to approximately 23% estimated by immunoblotting and supported by immunocytochemistry and by the amount of megalin recovered by RAP affinity chromatography. Light-and electron microscope immunocytochemistry as well as analyses on separated membrane fractions showed significant changes in the subcellular distribution of megalin. A significant reduction in the normal brush border labelling was observed in association with increased labeling of rough endoplasmic reticulum and the smooth paramembranous endoplasmic reticulum along the basolateral membranes. RAP deficiency was associated with changes in urinary protein composition, enabling the identification of α- amylase as a new ligand for megalin. In addition, an increased excretion of vitamin D-binding protein, a recently identified ligand to megalin, was observed Supporting changes in tubular protein reabsorption. The present data show that RAP is of crucial importance for normal processing and function of megalin, suggesting a chaperone-like function of this protein in the kidney proximal tubule.

AB - The receptor-associated protein (RAP) has been identified as a chaperone regulating the expression and processing of the LDL receptor-related protein. RAP also binds to the related 600-kD multiligand endocytic receptor megalin expressed in many absorptive epithelia including renal proximal tubule. The present study examines the effect of RAP gene disruption on megalin expression and subcellular distribution in the proximal tubule as well as the effect on tubular protein reabsorption. It is shown that RAP is important for the normal expression and function of megalin. Megalin expression was reduced to approximately 23% estimated by immunoblotting and supported by immunocytochemistry and by the amount of megalin recovered by RAP affinity chromatography. Light-and electron microscope immunocytochemistry as well as analyses on separated membrane fractions showed significant changes in the subcellular distribution of megalin. A significant reduction in the normal brush border labelling was observed in association with increased labeling of rough endoplasmic reticulum and the smooth paramembranous endoplasmic reticulum along the basolateral membranes. RAP deficiency was associated with changes in urinary protein composition, enabling the identification of α- amylase as a new ligand for megalin. In addition, an increased excretion of vitamin D-binding protein, a recently identified ligand to megalin, was observed Supporting changes in tubular protein reabsorption. The present data show that RAP is of crucial importance for normal processing and function of megalin, suggesting a chaperone-like function of this protein in the kidney proximal tubule.

U2 - 10.1681/ASN.V112191

DO - 10.1681/ASN.V112191

M3 - Journal article

C2 - 10665926

AN - SCOPUS:0033968875

SN - 1046-6673

VL - 11

SP - 191

EP - 202

JO - Journal of the American Society of Nephrology

JF - Journal of the American Society of Nephrology

IS - 2

ER -

Receptor-associated protein is important for normal processing of megalin in kidney proximal tubules

Abstract

Bibliografisk note

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