Linked Life Data

11468271

Source:http://linkedlifedata.com/resource/pubmed/id/11468271

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
15
pubmed:dateCreated
2001-7-24
pubmed:abstractText
PHEX is homologous to the M13 zinc metallopeptidases, a class of type II membrane glycoproteins. Although more than 140 mutations in the PHEX gene have been identified in patients with X-linked hypophosphatemia (XLH), the most prevalent form of inherited rickets, the molecular consequences of disease-causing PHEX mutations have not yet been investigated. We examined the effect of PHEX missense mutations on cellular trafficking of the recombinant protein. Four mutant PHEX cDNAs were generated by PCR mutagenesis: C85R, G579R and S711R, identified in XLH patients, and E581V, previously engineered in neutral endopeptidase 24.11, where it abolished catalytic activity but not plasma membrane targeting. Wild-type and mutant PHEX cDNAs were transfected in HEK(293) cells and PHEX protein expression was characterized. In contrast to the wild-type and E581V PHEX proteins, the C85R, G579R and S711R mutants were completely sensitive to endoglycosidase H digestion, indicating that they were not fully glycosylated. Sequestration of the disease-causing mutant proteins in the endoplasmic reticulum (ER) and plasma membrane localization of wild-type and E581V PHEX proteins was demonstrated by immunofluorescence and cell surface biotinylation. Of the three mutant PHEX proteins, the S711R was the least stable and the only one that could be rescued from the ER to the plasma membrane in cells grown at 26 degrees C. The chemical chaperone glycerol failed to correct defective targeting of all three mutant proteins. Our data provide a mechanism for loss of PHEX function in XLH patients expressing the C85R, G579R and S711R mutations.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0964-6906
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
10
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1539-46
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed-meshheading:11468271-Biotinylation, pubmed-meshheading:11468271-Calcium-Binding Proteins, pubmed-meshheading:11468271-Calnexin, pubmed-meshheading:11468271-Cell Line, pubmed-meshheading:11468271-Cell Membrane, pubmed-meshheading:11468271-DNA, Complementary, pubmed-meshheading:11468271-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:11468271-Exons, pubmed-meshheading:11468271-Genetic Linkage, pubmed-meshheading:11468271-Glycosylation, pubmed-meshheading:11468271-Hexosaminidases, pubmed-meshheading:11468271-Humans, pubmed-meshheading:11468271-Hypophosphatemia, Familial, pubmed-meshheading:11468271-Immunoblotting, pubmed-meshheading:11468271-Microscopy, Fluorescence, pubmed-meshheading:11468271-Mutation, pubmed-meshheading:11468271-Mutation, Missense, pubmed-meshheading:11468271-PHEX Phosphate Regulating Neutral Endopeptidase, pubmed-meshheading:11468271-Precipitin Tests, pubmed-meshheading:11468271-Protein Binding, pubmed-meshheading:11468271-Protein Transport, pubmed-meshheading:11468271-Proteins, pubmed-meshheading:11468271-Recombinant Proteins, pubmed-meshheading:11468271-Transfection, pubmed-meshheading:11468271-X Chromosome
pubmed:year
2001
pubmed:articleTitle
Disease-causing missense mutations in the PHEX gene interfere with membrane targeting of the recombinant protein.
pubmed:affiliation
Department of Biology, McGill University, Montreal, Quebec, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't