Department of Biomedical Sciences, University of Sassari, Sassari, Italy.
Methods Mol Biol. 2023;2619:239-248. doi: 10.1007/978-1-0716-2946-8_17.
This chapter describes a method for the purification of urinary trypsin inhibitor (UTI), a small chondroitin sulfate proteoglycan with Ser-proteinase inhibitory activity, excreted at high levels into urine following an inflammatory condition. The method consists of two fractionation steps: an anion-exchange chromatography and a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Coomassie Brilliant Blue G-250 gel staining. Several UTI bands are excised from gel, minced, destained, and dehydrated for extraction with SDS-containing buffer, at 60 °C for 24 h. This allows for obtaining a highly purified UTI sample useful for both structural and functional studies.
本章描述了一种从尿液中提取高浓度分泌型丝氨酸蛋白酶抑制剂（UTI）的方法。UTI 是一种小分子硫酸软骨素蛋白聚糖，具有丝氨酸蛋白酶抑制活性。该方法包括两步分离步骤：阴离子交换层析和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE），然后进行考马斯亮蓝 G-250 凝胶染色。从凝胶中切下几个 UTI 条带，切碎，脱色，并用含 SDS 的缓冲液在 60°C 下提取 24 小时进行脱水。这样可以获得高度纯化的 UTI 样品，可用于结构和功能研究。
