title: Selected reaction monitoring to differentiate and relatively quantitate isomers of sulfated and unsulfated core 1 O-glycans from salivary MUC7 protein in rheumatoid arthritis.
abstract: Rheumatoid arthritis is a common and debilitating systemic inflammatory condition affecting up to 1% of the world's population. This study aimed to investigate the immunological significance of O-glycans in chronic arthritis at a local and systemic level. O-Glycans released from synovial glycoproteins during acute and chronic arthritic conditions were compared and immune-reactive glycans identified. The sulfated core 1 O-glycan (Galβ1-3GalNAcol) was immune reactive, showing a different isomeric profile in the two conditions. From acute reactive arthritis, three isomers could be sequenced, but in patients with chronic rheumatoid arthritis, only a single 3-Gal sulfate-linked isomer could be identified. The systemic significance of this glycan epitope was investigated using the salivary mucin MUC7 in patients with rheumatoid arthritis and normal controls. To analyze this low abundance glycan, a selected reaction monitoring (SRM) method was developed to differentiate and relatively quantitate the core 1 O-glycan and the sulfated core 1 O-glycan Gal- and GalNAc-linked isomers. The acquisition of highly sensitive full scan linear ion trap MS/MS spectra in addition to quantitative SRM data allowed the 3- and 6-linked Gal isomers to be differentiated. The method was used to relatively quantitate the core 1 glycans from MUC7 to identify any systemic changes in this carbohydrate epitope. A statistically significant increase in sulfation was identified in salivary MUC7 from rheumatoid arthritis patients. This suggests a potential role for this epitope in chronic inflammation. This study was able to develop an SRM approach to specifically identify and relatively quantitate sulfated core 1 isomers and the unsulfated structure. The expansion of this method may afford an avenue for the high throughput investigation of O-glycans.
authors: Department of Medical Biochemistry, Institute of Biomedicine, University of Gothenburg, Medicinaregatan 9A, 405 30, Gothenburg, Sweden.
authors: Sarah A
authors: SA
authors: Flowers
authors: Liaqat
authors: L
authors: Ali
authors: Catherine S
authors: CS
authors: Lane
authors: Magnus
authors: M
authors: Olin
authors: Niclas G
authors: NG
authors: Karlsson
mesh_terms: D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006031:Glycosylation; D006595:Hexosamines / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007536:Isomerism; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012463:Saliva / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000378:metabolism*; D013463:Sulfuric Acid Esters / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D006595:Hexosamines; C083238:MUC7 protein, human; D009077:Mucins; D011134:Polysaccharides; D012471:Salivary Proteins and Peptides; D013463:Sulfuric Acid Esters