Uchimura Kenji
Department of Biochemistry, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi, 466-8550, Japan,
Methods Mol Biol. 2015;1229:401-12. doi: 10.1007/978-1-4939-1714-3_31.
Sulf-1 and Sulf-2 are endo-acting extracellular sulfatases. The Sulfs liberate 6-O sulfate groups, mainly from N, 6-O, and 2-O trisulfated disaccharides of heparan sulfate (HS)/heparin chains. The Sulfs have been shown to modulate the interaction of a number of protein ligands including growth factors and morphogens with HS/heparin and thus regulate the signaling of these ligands. They also play important roles in development and are dysregulated in many cancers. The establishment of the expression of the Sulfs and methods of assaying them has been desirable to investigate these enzymes. In this chapter, methods to express and purify recombinant Sulfs and to analyze HS structures in an extracellular fraction of HSulf-transfected HEK293 cells are described. The application of these enzymes for ex vivo degradation of an anti-HS epitope accumulated in the brain of a neurodegenerative disease model mouse is also described.
硫酸酯酶-1（Sulf-1）和硫酸酯酶-2（Sulf-2）是内源性细胞外硫酸酯酶。这些硫酸酯酶主要从硫酸乙酰肝素（HS）/肝素链的N、6-O和2-O三硫酸化二糖中释放6-O硫酸基团。已证明硫酸酯酶可调节包括生长因子和形态发生素在内的多种蛋白质配体与HS/肝素的相互作用，从而调节这些配体的信号传导。它们在发育过程中也发挥着重要作用，并且在许多癌症中表达失调。为了研究这些酶，建立硫酸酯酶的表达及检测方法很有必要。在本章中，将描述表达和纯化重组硫酸酯酶以及分析HSulf转染的HEK293细胞细胞外部分中HS结构的方法。还将描述这些酶在体外降解神经退行性疾病模型小鼠脑中积累的抗HS表位的应用。
