Zimmerhackl L B, Leuk B, Hoschützky H
Department of Pediatrics, University of Freiburg, Germany.
J Chromatogr. 1991 Nov 29;587(1):81-4. doi: 10.1016/0021-9673(91)85200-y.
Villin is a cytoskeletal protein of brush borders in the kidney and gut. After renal tubular cell injury the brushborder fragments are shedded into the tubular lumen and excreted with urine indicating renal tubular damage (so called "renal antigen" shedding). In urine villin appears as intact molecule (95,000 dalton) and as fragment with 70,000, 45,000 and 22,000 dalton. The major villin fragment (70,000 dalton) was purified after ammonium sulphate precipitation from urine of human renal transplant recipients. Final purification of the villin 70,000 dalton fragment was achieved by gel filtration with TSK 3000 SWG preparative grade. Purification was varified by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blotting.
绒毛蛋白是肾脏和肠道刷状缘的一种细胞骨架蛋白。肾小管细胞损伤后,刷状缘片段脱落至肾小管腔并随尿液排出,提示肾小管损伤(即所谓的“肾抗原”脱落)。在尿液中,绒毛蛋白以完整分子(95,000道尔顿)以及70,000、45,000和22,000道尔顿的片段形式出现。主要的绒毛蛋白片段(70,000道尔顿)是从人类肾移植受者的尿液中经硫酸铵沉淀后纯化得到的。通过使用TSK 3000 SWG制备级凝胶过滤对70,000道尔顿的绒毛蛋白片段进行最终纯化。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳和蛋白质印迹法对纯化进行验证。