Saklatvala J, Curry V A, Sarsfield S J
Biochem J. 1983 Nov 1;215(2):385-92. doi: 10.1042/bj2150385.
Catabolin, a protein that causes proteoglycan resorption in explants of living cartilage, was purified to homogeneity from culture medium conditioned by culturing buffy-coat leucocytes from 60 litres of pig blood in the presence of concanavalin A. The purification steps were (1) gel filtration of concentrated medium, (2) chromatofocusing, (3) hydroxyapatite chromatography, (4) anion-exchange chromatography (Mono Q), (5) reversed-phase high-pressure liquid chromatography (h.p.l.c.) (Zorbax ODS). These achieved approx. 9000-fold purification from the starting material. The purified protein when reduced ran as a single band on sodium dodecyl sulphate (SDS)/polyacrylamide-gel electrophoresis with Mr 21000. On isoelectric focusing its pI was 4.8-5.0, and there was evidence of micro-heterogeneity. The protein co-migrated with active material on h.p.l.c., isoelectric focusing and SDS gels (15 and 12.5% acrylamide) under both reducing and non-reducing conditions. The pure protein caused proteoglycan release from cultured bovine nasal cartilage at 20pM. Its possible identity with interleukin 1 is discussed.
分解代谢素是一种能导致活软骨外植体中蛋白聚糖重吸收的蛋白质，它是从在伴刀豆球蛋白A存在的情况下培养60升猪血中的血沉棕黄层白细胞所制备的条件培养基中纯化至同质的。纯化步骤如下：（1）浓缩培养基的凝胶过滤；（2）色谱聚焦；（3）羟基磷灰石色谱法；（4）阴离子交换色谱法（Mono Q）；（5）反相高压液相色谱法（h.p.l.c.）（Zorbax ODS）。这些步骤使起始原料实现了约9000倍的纯化。纯化后的蛋白质在还原条件下于十二烷基硫酸钠（SDS）/聚丙烯酰胺凝胶电泳上呈现为一条分子量为21000的单带。在等电聚焦中，其pI为4.8 - 5.0，并且有微异质性的证据。该蛋白质在还原和非还原条件下，在h.p.l.c.、等电聚焦和SDS凝胶（15%和12.5%丙烯酰胺）上均与活性物质共迁移。纯蛋白质在20pM时可导致培养的牛鼻软骨释放蛋白聚糖。文中讨论了它与白细胞介素1可能的同一性。
