Molecular cloning of the major cell surface heparan sulfate proteoglycan from rat liver.

2.50
Hdl Handle:
http://hdl.handle.net/10541/108944
Title:
Molecular cloning of the major cell surface heparan sulfate proteoglycan from rat liver.
Authors:
Pierce, Andrew; Lyon, Malcolm; Hampson, Ian N; Cowling, Graham J; Gallagher, John T
Abstract:
We have used an antiserum raised against a purified heparan sulfate proteoglycan (HSPG) preparation isolated from rat liver to screen a lambda gt11 expression library and have obtained overlapping cDNA clones that contain the full-length coding sequence of an HSPG core protein capable of spanning the plasma membrane. The open reading frame of the rat cDNA encodes a protein of 211 amino acids. The predicted protein sequence (23 kDa) has a high degree of homology with the published partial sequence of the human lung fibroblast HSPG, fibroglycan. The deduced protein sequence contains a 24-amino acid transmembrane domain and a 33-amino acid cytoplasmic domain, both of which are identical with the corresponding regions of human fibroglycan and are highly homologous to the human, hamster, and mouse epithelial HSPG, syndecan. The putative ectodomain, which has 85% homology to fibroglycan, contains three possible glycosaminoglycan attachment sites that may be occupied by heparan sulfate chains. The major 49-kDa core protein in the liver HSPG preparation was found to be reactive to an antibody that specifically recognizes the cytoplasmic domain of fibroglycan. We have used the full-length cDNA clone to analyze the expression of this transmembrane core protein gene in whole tissues and several epithelial and fibroblastoid cell lines. It hybridizes to three mRNA species in all cell and tissue types examined, but in liver, isolated hepatocytes, and kidney, an additional 0.8-kilobase mRNA was detected. The three common messages arise from differential use of alternative polyadenylation sites, whereas the fourth tissue-restricted RNA species represents a related gene transcript. The rat equivalent of human fibroglycan therefore appears to be the major transmembrane proteoglycan in liver, and its widespread expression in many diverse tissues and cells suggests that it plays an important role in cellular interactions.
Affiliation:
Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom.
Citation:
Molecular cloning of the major cell surface heparan sulfate proteoglycan from rat liver. 1992, 267 (6):3894-900 J. Biol. Chem.
Journal:
Journal of Biological Chemistry
Issue Date:
25-Feb-1992
URI:
http://hdl.handle.net/10541/108944
PubMed ID:
1740437
Type:
Article
Language:
en
ISSN:
0021-9258
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorPierce, Andrewen
dc.contributor.authorLyon, Malcolmen
dc.contributor.authorHampson, Ian Nen
dc.contributor.authorCowling, Graham Jen
dc.contributor.authorGallagher, John Ten
dc.date.accessioned2010-08-03T11:34:05Z-
dc.date.available2010-08-03T11:34:05Z-
dc.date.issued1992-02-25-
dc.identifier.citationMolecular cloning of the major cell surface heparan sulfate proteoglycan from rat liver. 1992, 267 (6):3894-900 J. Biol. Chem.en
dc.identifier.issn0021-9258-
dc.identifier.pmid1740437-
dc.identifier.urihttp://hdl.handle.net/10541/108944-
dc.description.abstractWe have used an antiserum raised against a purified heparan sulfate proteoglycan (HSPG) preparation isolated from rat liver to screen a lambda gt11 expression library and have obtained overlapping cDNA clones that contain the full-length coding sequence of an HSPG core protein capable of spanning the plasma membrane. The open reading frame of the rat cDNA encodes a protein of 211 amino acids. The predicted protein sequence (23 kDa) has a high degree of homology with the published partial sequence of the human lung fibroblast HSPG, fibroglycan. The deduced protein sequence contains a 24-amino acid transmembrane domain and a 33-amino acid cytoplasmic domain, both of which are identical with the corresponding regions of human fibroglycan and are highly homologous to the human, hamster, and mouse epithelial HSPG, syndecan. The putative ectodomain, which has 85% homology to fibroglycan, contains three possible glycosaminoglycan attachment sites that may be occupied by heparan sulfate chains. The major 49-kDa core protein in the liver HSPG preparation was found to be reactive to an antibody that specifically recognizes the cytoplasmic domain of fibroglycan. We have used the full-length cDNA clone to analyze the expression of this transmembrane core protein gene in whole tissues and several epithelial and fibroblastoid cell lines. It hybridizes to three mRNA species in all cell and tissue types examined, but in liver, isolated hepatocytes, and kidney, an additional 0.8-kilobase mRNA was detected. The three common messages arise from differential use of alternative polyadenylation sites, whereas the fourth tissue-restricted RNA species represents a related gene transcript. The rat equivalent of human fibroglycan therefore appears to be the major transmembrane proteoglycan in liver, and its widespread expression in many diverse tissues and cells suggests that it plays an important role in cellular interactions.en
dc.language.isoenen
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshBlotting, Northern-
dc.subject.meshBlotting, Southern-
dc.subject.meshBlotting, Western-
dc.subject.meshCell Membrane-
dc.subject.meshCloning, Molecular-
dc.subject.meshDNA-
dc.subject.meshElectrophoresis, Polyacrylamide Gel-
dc.subject.meshFibroblasts-
dc.subject.meshHeparan Sulfate Proteoglycans-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshLiver-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshNucleic Acid Hybridization-
dc.subject.meshPrecipitin Tests-
dc.subject.meshProteoglycans-
dc.subject.meshRNA, Messenger-
dc.subject.meshRats-
dc.titleMolecular cloning of the major cell surface heparan sulfate proteoglycan from rat liver.en
dc.typeArticleen
dc.contributor.departmentDepartment of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom.en
dc.identifier.journalJournal of Biological Chemistryen
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