Bhatt A K, Spilberg I
Veterans Administration Medical Center, Rheumatology Unit, St. Louis, MO 63106.
Clin Biochem. 1988 Dec;21(6):341-5. doi: 10.1016/s0009-9120(88)80014-6.
The crystal induced chemotactic factor is a protein that is released by neutrophils in response to a phagocytic stimulus like monosodium urate or calcium pyrophosphate crystals. This protein has been postulated to play an important role in the development of crystal-induced arthritis. In this report, we present a protocol for its purification which involved ammonium sulfate fractionation, affinity chromatography, lectin chromatography, and high pressure chromatography (HPLC). The purified protein migrated as a single band on SDS-PAGE. Based on its electrophoretic and chromatographic (HPLC) properties, it appears to have a m.w. of 15,000. It has a blocked N-terminal since our attempt to sequence the chemotactic protein by established procedures failed. The protein contains one methionine residue which appears to be essential for its chemotactic activity since cyanogen bromide treatment abolished its chemotactic activity. To our knowledge this is the first report on purification of this autocrine leukoattractant from human neutrophils.
晶体诱导趋化因子是一种蛋白质，由中性粒细胞在受到吞噬刺激（如尿酸钠或焦磷酸钙晶体）时释放。据推测，这种蛋白质在晶体诱导性关节炎的发展中起重要作用。在本报告中，我们展示了一种其纯化方案，该方案包括硫酸铵分级分离、亲和色谱、凝集素色谱和高压色谱（HPLC）。纯化后的蛋白质在SDS-PAGE上呈单一泳带迁移。根据其电泳和色谱（HPLC）特性，其分子量似乎为15,000。由于我们尝试用既定方法对趋化蛋白进行测序失败，其N端被封闭。该蛋白质含有一个甲硫氨酸残基，由于溴化氰处理消除了其趋化活性，该残基似乎对其趋化活性至关重要。据我们所知，这是关于从人中性粒细胞中纯化这种自分泌白细胞趋化因子的首次报告。
