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Compositional profiling of heparin/heparan sulfate using mass spectrometry: assay for specificity of a novel extracellular human endosulfatase.
- Source :
-
Glycobiology [Glycobiology] 2005 Aug; Vol. 15 (8), pp. 818-26. Date of Electronic Publication: 2005 Apr 20. - Publication Year :
- 2005
-
Abstract
- An important class of carbohydrates studied within the field of glycobiology, heparin and heparan sulfate (HS) have been implicated in a diverse array of biological functions. Changes in their sulfation pattern and domain organization have been associated with different pathological situations such as viral infectivity, tumor growth, and metastasis. To obtain structural information about these biomolecules, and the modifications they may undergo during different stages of cell growth and development, a mass spectrometry-based method was developed and used to obtain unambiguous structural information on the glycosaminoglycans (GAGs) that comprise heparin/HS. The method was applied to assay for the heparin substrate specificity of a newly discovered human extracellular endosulfatase, HSulf-2, which has been implicated in tumorigenesis. This new protocol incorporates 12 known heparin disaccharides, including three sets of isomers. A unique response factor (R) is determined for each disaccharide, whereas a multiplexed and data processing method is incorporated for faster data acquisition and quantification purposes. Proof of principle was performed by using various heparin/HS samples isolated from bovine and porcine tissues.
- Subjects :
- Animals
Breast Neoplasms enzymology
Cattle
Heparin analysis
Humans
Intestinal Mucosa metabolism
Kidney metabolism
Lung metabolism
Protein Binding
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization
Spectrometry, Mass, Fast Atom Bombardment
Sulfatases isolation & purification
Swine
Biological Assay
Disaccharides analysis
Heparin analogs & derivatives
Heparitin Sulfate chemistry
Sulfatases metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0959-6658
- Volume :
- 15
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Glycobiology
- Publication Type :
- Academic Journal
- Accession number :
- 15843596
- Full Text :
- https://doi.org/10.1093/glycob/cwi064