Bumol T F, Walker L E, Reisfeld R A
J Biol Chem. 1984 Oct 25;259(20):12733-41.
Monoclonal antibody (Mab) 9.2.27 was utilized in a combination of biosynthetic and biochemical investigations as an immunological probe for the study of chondroitin sulfate proteoglycans (CSP) in human melanoma cells. Pulse-chase and long-term intrinsic labeling immunoprecipitation experiments combined with the biosynthetic inhibitors monensin, cycloheximide, and paranitrophenol-beta-D-xyloside all suggest that Mab 9.2.27 recognizes a set of glycoprotein molecules ranging to a 250-kDa glycoprotein which serves as the core glycoprotein for CSP in human melanoma cells. Peptide maps comparing the 250-kDa and CSP molecule verify that the 250-kDa glycoprotein is the CSP core protein in human melanoma cells. Further studies document that the CSP released by melanoma cells and recognized by Mab 9.2.27 contains (2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-beta-4-O-sulfo-D-galactose and 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-beta-6-O-sulfo-D-galactose saccharides and this CSP can interact with hyaluronic acid-Sepharose. Topographical studies indicate that this CSP has pericellular punctuated distribution on the melanoma cell surface and may play a role in cell-substrate interactions in the biology of metastatic human melanoma.
单克隆抗体（Mab）9.2.27被用于生物合成和生化研究的组合中，作为一种免疫探针，用于研究人黑色素瘤细胞中的硫酸软骨素蛋白聚糖（CSP）。脉冲追踪和长期内在标记免疫沉淀实验，结合生物合成抑制剂莫能菌素、环己酰亚胺和对硝基苯酚-β-D-木糖苷，均表明Mab 9.2.27识别一组糖蛋白分子，范围至一种250 kDa的糖蛋白，该糖蛋白是人黑色素瘤细胞中CSP的核心糖蛋白。比较250 kDa和CSP分子的肽图证实，250 kDa糖蛋白是人黑色素瘤细胞中的CSP核心蛋白。进一步研究表明，黑色素瘤细胞释放并被Mab 9.2.27识别的CSP含有（2-乙酰氨基-2-脱氧-3-O-（β-D-葡萄糖-4-烯吡喃糖醛酸）-β-4-O-磺基-D-半乳糖和2-乙酰氨基-2-脱氧-3-O-（β-D-葡萄糖-4-烯吡喃糖醛酸）-β-6-O-磺基-D-半乳糖糖类，并且这种CSP可以与透明质酸-琼脂糖相互作用。拓扑学研究表明，这种CSP在黑色素瘤细胞表面具有细胞周围点状分布，可能在转移性人黑色素瘤生物学中的细胞-底物相互作用中发挥作用。
