Hitchcock Alicia M, Yates Karen E, Costello Catherine E, Zaia Joseph
Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
Proteomics. 2008 Apr;8(7):1384-97. doi: 10.1002/pmic.200700787.
Homeostasis of connective joint tissues depends on the maintenance of an extracellular matrix, consisting of an integrated assembly of collagens, glycoproteins, proteoglycans, and glycosaminoglycans (GAGs). Isomeric chondroitin sulfate (CS) glycoforms differing in position and degree of sulfation and uronic acid epimerization play specific and distinct functional roles during development and disease onset. This work profiles the CS epitopes expressed by different joint tissues as a function of age and osteoarthritis. GAGs were extracted from joint tissues (cartilage, tendon, ligment, muscle, and synovium) and partially depolymerized using chondroitinase enzymes. The oligosaccharide products were differentially stable isotope labeled by reductive amination using 2-anthranilic acid-d(0) or -d(4) and subjected to amide-hydrophilic interaction chromatography (HILIC) online LC-MS/MS. The analysis presented herein enables simultaneous profiling of the expression of nonreducing end, linker region, and Delta-unsaturated interior oligosaccharide domains of the CS chains among the different joint tissues. The results provide important new information on the changes to the expression of CS GAG chains during disease and development.
结缔关节组织的稳态取决于细胞外基质的维持，该基质由胶原蛋白、糖蛋白、蛋白聚糖和糖胺聚糖（GAGs）的整合组装体组成。硫酸软骨素（CS）糖异构体在硫酸化位置和程度以及糖醛酸差向异构化方面存在差异，在发育和疾病发生过程中发挥着特定且独特的功能作用。这项工作描绘了不同关节组织所表达的CS表位随年龄和骨关节炎的变化情况。从关节组织（软骨、肌腱、韧带、肌肉和滑膜）中提取GAGs，并使用软骨素酶进行部分解聚。通过用2-氨基苯甲酸-d(0)或-d(4)进行还原胺化对寡糖产物进行差异稳定同位素标记，然后进行酰胺-亲水相互作用色谱（HILIC）在线LC-MS/MS分析。本文所呈现的分析能够同时描绘不同关节组织中CS链的非还原端、连接区和Δ-不饱和内部寡糖结构域的表达情况。这些结果为疾病和发育过程中CS GAG链表达的变化提供了重要的新信息。
