RGD Reference Report - Interactions of cubilin with megalin and the product of the amnionless gene (AMN): effect on its stability. - Rat Genome Database

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Interactions of cubilin with megalin and the product of the amnionless gene (AMN): effect on its stability.

Authors: Ahuja, R  Yammani, R  Bauer, JA  Kalra, S  Seetharam, S  Seetharam, B 
Citation: Ahuja R, etal., Biochem J. 2008 Mar 1;410(2):301-8.
RGD ID: 2317831
Pubmed: PMID:17990981   (View Abstract at PubMed)
DOI: DOI:10.1042/BJ20070919   (Journal Full-text)

Cubilin, a 456 kDa multipurpose receptor lacking in both transmembrane and cytoplasmic domains is expressed in the apical BBMs (brush border membranes) of polarized epithelia. Cubilin interacts with two transmembrane proteins, AMN, a 45-50 kDa protein product of the amnionless gene, and megalin, a 600 kDa giant endocytic receptor. In vitro, three fragments of cubilin, the 113-residue N-terminus and CUB domains 12-17 and 22-27, demonstrated Ca2+-dependent binding to megalin. Immunoprecipitation and immunoblotting studies using detergent extracts of rat kidney BBMs revealed that cubilin interacts with both megalin and AMN. Ligand (intrinsic factor-cobalamin)-affinity chromatography showed that in renal BBMs, functional cubilin exists as a complex with both AMN and megalin. Cubilin and AMN levels were reduced by 80% and 55-60% respectively in total membranes and BBMs obtained from kidney of megalin antibody-producing rabbits. Immunohistochemical analysis and turnover studies for cubilin in megalin or AMN gene-silenced opossum kidney cells showed a significant reduction (85-90%) in cubilin staining and a 2-fold decrease in its half-life. Taken together, these results indicate that three distinct regions of cubilin bind to megalin and its interactions with both megalin and AMN are essential for its intracellular stability.

Gene Ontology Annotations    Click to see Annotation Detail View

Cellular Component
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
brush border membrane  IDA 2317831; 2317831 RGD 
protein-containing complex  IDA 2317831; 2317831 RGD 

Objects Annotated

Genes (Rattus norvegicus)
Amn  (amnion associated transmembrane protein)
Cubn  (cubilin)


Additional Information