Studies on the Extraction of Different Proteoglycan Populations in Bovine Articular Cartilage

Hs, Brand; Korver, G. H. V.; Stadt, R J van de; Kampen, G P van; Korst, J. K. van der

doi:10.1515/bchm3.1990.371.2.581

Cited by 6 publications

(1 citation statement)

References 14 publications

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“…4M GuHCl has proven to offer the best extraction yield and has become the most widely used extraction agent. Brand et al [33] extracted bovine articular cartilage with increasing concentrations of GuHCl and concluded that the ionic strength employed for the efficient solubilization of PGs is directly proportional to their size. However, for tissues less abundant in proteoglycans, GuHCl extraction results also in a high amount of other proteins.…”

Section: Extraction From Tissuementioning

confidence: 99%

Recent Developments in Proteoglycan Purification and Analysis

Didraga¹,

Barroso²,

Bischoff³

2006

CPA

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Proteoglycans are ubiquitous biomolecules in the body located in the extracellular matrix, on the cell surface and also within the cells. They contain at least one glycosaminoglycan (GAG) chain covalently attached to a core protein and may also present N-or O-linked glycans. The high structural diversity and distribution relate to the various biological functions of proteoglycans. In recent years, new members have enlarged the proteoglycan family and advances in molecular biology and glycobiology contributed to elucidate more of the biological functions of proteoglycans. In order to study the structure of a proteoglycan molecule and relate it to its function (or dysfunction), its isolation and purification from cell culture or tissue extracts is necessary. Next to the widely used anion exchange chromatographic methods, techniques based on lectin affinity chromatography have created new possibilities to increase the degree of purity. Introduction of electrospray ionization (ESI) and matrix-assisted laser desorption-ionization (MALDI) sources, together with tandem mass spectrometry (MS/MS or MS n ), mark a further important step towards the structural analysis of glycosaminoglycans. The aim of this review is to present the most recent advances in proteoglycan purification and analysis.

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Section: Extraction From Tissuementioning

confidence: 99%

Recent Developments in Proteoglycan Purification and Analysis

Didraga¹,

Barroso²,

Bischoff³

2006

CPA

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Isolation and purification of proteoglycans

Fedarko

1994

Proteoglycans

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Isolation and purification of proteoglycans

Fedarko

1993

Experientia

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Purification of a protein typically involves development of a quantitative assay to track protein integrity (e.g. enzyme activity) during subsequent isolation steps. The generalized procedure involves choosing the source of the protein, defining extraction conditions, developing bulk purification methods followed by refined, more selective methods. The purification of proteoglycans is often complicated by a) limited source quantities, b) necessity of chaotropic solvents for efficient extraction, c) their large molecular size and d) lack of defined functions to enable purity (i.e. activity, conformation) to be assessed. Because the usual goal of proteoglycan purification is physical characterization (intact molecular weight, core protein and glycosaminoglycan class and size), the problems of a suitable assay and/or native conformation are avoided. The 'assay' for tracking proteoglycan isolation typically utilizes uronic acid content or radiolabel incorporation as a marker. Once extracted from their cellular/extracellular environment, proteoglycans can be isolated by density gradient centrifugation and/or column chromatography techniques. Recent advances in the composition of chromatographic supports have enabled the application of ion-exchange, gel permeation, hydrophobic interaction and affinity chromatography resins using efficient high-pressure liquid chromatography to proteoglycan purification.

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Studies on the Extraction of Different Proteoglycan Populations in Bovine Articular Cartilage

Cited by 6 publications

References 14 publications

Recent Developments in Proteoglycan Purification and Analysis

Recent Developments in Proteoglycan Purification and Analysis

Isolation and purification of proteoglycans

Isolation and purification of proteoglycans

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