title: Detection of specific glycosaminoglycans and glycan epitopes by in vitro sulfation using recombinant sulfotransferases.
abstract: Sulfated glycans play critical roles during the development, differentiation and growth of various organisms. The most well-studied sulfated molecules are sulfated glycosaminoglycans (GAGs). Recent incidents of heparin drug contamination convey the importance of having a convenient and sensitive method for detecting different GAGs. Here, we describe a molecular method to detect GAGs in biological and biomedical samples. Because the sulfation of GAGs is generally not saturated in vivo, it is possible to introduce the radioisotope (35)S in vitro using recombinant sulfotransferases, thereby allowing detection of minute quantities of these molecules. This strategy was also successfully applied in the detection of other glycans. As examples, we detected contaminant GAGs in commercial heparin, heparan sulfate and chondroitin samples. The identities of the contaminant GAGs were further confirmed by lyase digestion. Oversulfated chondroitin sulfate was detectable only following a simple desulfation step. Additionally, in vitro sulfation by sulfotransferases allowed us to map glycan epitopes in biological samples. This was illustrated using mouse embryo and rat organ tissue sections labeled with the following carbohydrate sulfotransferases: CHST3, CHST15, HS3ST1, CHST4 and CHST10.
authors.affiliation: R&D Systems Inc., Minneapolis, MN 55413, USA. leon.wu@rndsystems.com
authors.forename: Zhengliang L
authors.initials: ZL
authors.lastname: Wu
authors.forename: Brittany
authors.initials: B
authors.lastname: Prather
authors.forename: Cheryl M
authors.initials: CM
authors.lastname: Ethen
authors.forename: Alex
authors.initials: A
authors.lastname: Kalyuzhny
authors.forename: Weiping
authors.initials: W
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D001923:Brain Chemistry; D002807:Chondroitin / Q000032:analysis; D019587:Dietary Supplements / Q000032:analysis; D004340:Drug Contamination / Q000517:prevention & control; D004622:Embryo, Mammalian / Q000737:chemistry / Q000378:metabolism; D005944:Glucosamine / Q000032:analysis; D006025:Glycosaminoglycans / Q000737:chemistry* / Q000378:metabolism; D006493:Heparin / Q000737:chemistry; D007668:Kidney / Q000737:chemistry / Q000378:metabolism; D008168:Lung / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D009130:Muscle, Smooth / Q000737:chemistry / Q000378:metabolism; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry*; D013194:Staining and Labeling; D015238:Sulfotransferases / Q000096:biosynthesis / Q000737:chemistry*; D013462:Sulfur Radioisotopes
chemical_list: D006025:Glycosaminoglycans; D011994:Recombinant Proteins; D013462:Sulfur Radioisotopes; D006493:Heparin; D002807:Chondroitin; D015238:Sulfotransferases; D005944:Glucosamine