Isolation and characterization of the integral glycosaminoglycan constituents of human amyloid A and monoclonal light-chain amyloid fibrils.

2.50
Hdl Handle:
http://hdl.handle.net/10541/106578
Title:
Isolation and characterization of the integral glycosaminoglycan constituents of human amyloid A and monoclonal light-chain amyloid fibrils.
Authors:
Nelson, S R; Lyon, Malcolm; Gallagher, John T; Johnson, E A; Pepys, M B
Abstract:
Amyloid fibrils were isolated by extraction in water from the livers and spleens of four patients who had died of monoclonal, light-chain (AL)-type, systemic amyloidosis and one with reactive systemic, amyloid A protein (AA)-type amyloidosis. Each fibril preparation contained 1-2% by weight of glycosaminoglycan (GAG) which was tightly associated with the fibrils and not just co-isolated from the tissues with them. After exhaustive digestion of the fibrils with papain and Pronase, the GAGs were specifically precipitated with cetylpyridinium chloride and were identified by cellulose acetate electrophoresis and selective susceptibility to specific glycosidases. All the preparations contained approximately equal amounts of heparan sulphate and dermatan sulphate. There was no evidence for the presence of chondroitin sulphate or other GAGs. Fine structural analysis by oligosaccharide mapping in gradient polyacrylamide gels, following partial digestion with specific glycosidases, showed very similar structures among the heparan sulphates and the dermatan sulphates, respectively. GAGs were also extracted by solubilizing amyloid fibrils in 4 M-guanidinium chloride followed by CsCl density-gradient ultracentrifugation. Although a minor proportion of the GAG material obtained in this way was apparently in the form of proteoglycan molecules, most of it was free GAG chains. The presence in amyloid fibrils of different types, in different organs and from different patients of particular GAG classes with similar structures supports the view that these molecules may be of pathogenic significance.
Affiliation:
Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.
Citation:
Isolation and characterization of the integral glycosaminoglycan constituents of human amyloid A and monoclonal light-chain amyloid fibrils. 1991, 275 ( Pt 1):67-73 Biochem. J.
Journal:
Biochemical Journal
Issue Date:
1-Apr-1991
URI:
http://hdl.handle.net/10541/106578
PubMed ID:
1902087
Type:
Article
Language:
en
ISSN:
0264-6021
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorNelson, S Ren
dc.contributor.authorLyon, Malcolmen
dc.contributor.authorGallagher, John Ten
dc.contributor.authorJohnson, E Aen
dc.contributor.authorPepys, M Ben
dc.date.accessioned2010-06-21T14:01:47Z-
dc.date.available2010-06-21T14:01:47Z-
dc.date.issued1991-04-01-
dc.identifier.citationIsolation and characterization of the integral glycosaminoglycan constituents of human amyloid A and monoclonal light-chain amyloid fibrils. 1991, 275 ( Pt 1):67-73 Biochem. J.en
dc.identifier.issn0264-6021-
dc.identifier.pmid1902087-
dc.identifier.urihttp://hdl.handle.net/10541/106578-
dc.description.abstractAmyloid fibrils were isolated by extraction in water from the livers and spleens of four patients who had died of monoclonal, light-chain (AL)-type, systemic amyloidosis and one with reactive systemic, amyloid A protein (AA)-type amyloidosis. Each fibril preparation contained 1-2% by weight of glycosaminoglycan (GAG) which was tightly associated with the fibrils and not just co-isolated from the tissues with them. After exhaustive digestion of the fibrils with papain and Pronase, the GAGs were specifically precipitated with cetylpyridinium chloride and were identified by cellulose acetate electrophoresis and selective susceptibility to specific glycosidases. All the preparations contained approximately equal amounts of heparan sulphate and dermatan sulphate. There was no evidence for the presence of chondroitin sulphate or other GAGs. Fine structural analysis by oligosaccharide mapping in gradient polyacrylamide gels, following partial digestion with specific glycosidases, showed very similar structures among the heparan sulphates and the dermatan sulphates, respectively. GAGs were also extracted by solubilizing amyloid fibrils in 4 M-guanidinium chloride followed by CsCl density-gradient ultracentrifugation. Although a minor proportion of the GAG material obtained in this way was apparently in the form of proteoglycan molecules, most of it was free GAG chains. The presence in amyloid fibrils of different types, in different organs and from different patients of particular GAG classes with similar structures supports the view that these molecules may be of pathogenic significance.en
dc.language.isoenen
dc.subject.meshAmyloidosis-
dc.subject.meshCarbohydrate Sequence-
dc.subject.meshCentrifugation, Density Gradient-
dc.subject.meshChemical Precipitation-
dc.subject.meshDermatan Sulfate-
dc.subject.meshElectrophoresis, Cellulose Acetate-
dc.subject.meshGlycosaminoglycans-
dc.subject.meshGuanidine-
dc.subject.meshGuanidines-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshHumans-
dc.subject.meshImmunoglobulin Light Chains-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshPapain-
dc.subject.meshPronase-
dc.subject.meshSerum Amyloid A Protein-
dc.titleIsolation and characterization of the integral glycosaminoglycan constituents of human amyloid A and monoclonal light-chain amyloid fibrils.en
dc.typeArticleen
dc.contributor.departmentDepartment of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.en
dc.identifier.journalBiochemical Journalen

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